{"title":"IHC \u0026 IF Reagents","description":"\u003ch5\u003e\n\u003cmeta charset=\"utf-8\"\u003e \u003cmeta charset=\"utf-8\"\u003e \u003cspan\u003e\u003cstrong\u003eBioss empowers your immunohistochemistry (IHC) and immunofluorescence (IF) research with a rigorously curated portfolio of high-specific antibodies and advanced detection reagents. Engineered to optimize staining precision and reproducibility, our solutions ensure confidence in your results.\u003c\/strong\u003e \u003c\/span\u003e\u003cbr\u003e\n\u003c\/h5\u003e","products":[{"product_id":"ihct003","title":"Multiplex Fluorescent (5 colors) IHC Staining Kit","description":"\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch1 style=\"text-align: center; margin: 0 0 10px 0; line-height: 1.1; background: transparent !important;\"\u003eMultiplex Fluorescent (5 colors) IHC Staining Kit\u003c\/h1\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eCat. No.:\u003c\/strong\u003e IHCT003\u003cbr\u003e\u003cstrong\u003eDetection method:\u003c\/strong\u003e Fluorescent\u003cbr\u003e\u003cstrong\u003eSection type:\u003c\/strong\u003e FFPE \u0026amp; Frozen Tissue, cells\u003cbr\u003e\u003cstrong\u003eSize:\u003c\/strong\u003e 20T | 100T\u003cbr\u003e\u003cstrong\u003eStorage and Stability:\u003c\/strong\u003e Store protected from light at 2–8℃ for 12 months.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Product Components --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch3 style=\"margin: 0 0 8px 0; line-height: 1.1; background: transparent !important;\"\u003eProduct Components\u003c\/h3\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; text-align: left; table-layout: fixed; line-height: 1.1; margin: 0 0 8px 0; background: transparent !important;\" class=\"product-component\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 28.0497%;\"\u003e \u003ccol style=\"width: 41.9804%;\"\u003e \u003ccol style=\"width: 14.872%;\"\u003e \u003ccol style=\"width: 15.0602%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent \u0026lt;p style=;\" rowspan=\"2\"\u003eName\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; background: transparent style=;\" rowspan=\"2\"\u003eComponent\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent \u0026lt;p style=;\" colspan=\"2\"\u003eSpecification\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003e20 T\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003e100 T\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e \u003cbr\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eNuclear dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eDAPI (Ready-to-use)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA Buffer\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e6.4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e32mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eHRP Goat Anti-Rabbit\/Mouse Universal Secondary Antibody\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e6.4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e32mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eAntibody Diluent\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e3% Hydrogen Peroxide\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eAnti-fade Mounting Medium\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTSA fluorophore working solution = TSA concentrated fluorophore + TSA buffer.\u003cbr\u003eThe dilution ratio should be optimized according to the specific experiment. The recommended range is 1:50 to 1:400, and 1:200 gives the best results in most cases.\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated at room temperature for 1–3 hours, a dye dilution of 1:50–1:200 is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated overnight at 4°C, a dilution of 1:200–1:400 or higher is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the signal is too strong, reduce the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eIf the signal is too weak, increase the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Principle --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRINCIPLE\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTyramide Signal Amplification (TSA) is an enzymatic detection method that uses horseradish peroxidase (HRP) to label target proteins in situ. The principle is based on the peroxidase reaction of tyramide. Under the action of HRP and H₂O₂, the tyramide fluorophore substrate becomes activated. The activated fluorophore then covalently binds to tyrosine and other residues on the target protein, causing substantial fluorophore deposition at the antigen–antibody binding site and thereby amplifying the signal.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter each staining round, the previously bound non-covalent antibodies can be removed by heat-induced retrieval or an antibody elution buffer, while the fluorophore remains stably attached to the protein. This enables the next round of staining. Once all antibody incubations are completed, nuclear staining, mounting, and scanning are performed.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eBecause only one antibody system is present in each staining round, there is no need to worry about antibody cross-reactivity or host-species matching between primary and secondary antibodies. This overcomes the species-origin limitations common in traditional immunofluorescence experiments.\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit may be used individually or in combination and can support single staining, double staining, triple staining, and even more multiplex fluorescence labeling.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Experimental Procedure --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eEXPERIMENTAL PROCEDURE\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSample Preparation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e1) Paraffin sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace slides sequentially into:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene I, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene II, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol I, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol II, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e95% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e85% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e75% ethanol, 5 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen rinse with distilled water.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e2) Frozen sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFix frozen sections for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e3) Cell climbing slides or cell smears\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFix cell samples for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntigen Retrieval\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace tissue sections into a retrieval container filled with either:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003epH 9.0 EDTA alkaline antigen retrieval solution, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003epH 6.0 citrate retrieval buffer\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePerform antigen retrieval in a microwave oven (other methods such as pressure cooker 1–2 min, boiling water at 100°C for 15 min, or water bath at 95°C for 20 min may also be used).\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eMicrowave program:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emedium heat for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003estop heating and stay still for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emedium-low heat for 7 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eDuring this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter natural cooling, place slides into PBS (pH 7.4), and wash on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe choice of retrieval buffer and conditions depends on tissue type, fixation method, and antigen type. This step may be omitted for frozen sections and cell samples.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Endogenous Peroxidase\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eIncubate the slides in 3% H₂O₂ solution at room temperature, protected from light, for 15 min. Then wash in PBS (pH 7.4) on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Non-specific Binding\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter gently removing excess liquid, draw a hydrophobic barrier around the tissue with a PAP pen.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd antibody diluent (or another blocking reagent such as 3% BSA or goat serum) to completely cover the tissue and block at room temperature for 30 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The antibody diluent contains various stabilizers and preservatives and can be used for blocking or for dilution of primary antibodies. Diluted primary antibodies can be stored long-term at 4°C and for up to one month at room temperature.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003ePrimary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eGently remove the blocking solution and add the appropriately diluted primary antibody working solution to the slide. Incubate flat in a humidified chamber protected from light:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eovernight at 4°C, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e1–2 h at 37°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd a small amount of water into the humid chamber to prevent antibody evaporation.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecondary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add the HRP-conjugated secondary antibody corresponding to the host species of the primary antibody to fully cover the tissue. Incubate at room temperature for 50 min, protected from light, then wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The kit includes a ready-to-use HRP goat anti-rabbit\/mouse universal secondary antibody with very high sensitivity. No preparation is required.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eFluorescence Staining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd the diluted TSA fluorophore working solution to completely cover the tissue and incubate at room temperature for 1–15 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended reaction time: 5–10 min\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e In a pilot experiment, stain for 1 min first, wash, and observe the result. If the positive signal is weak, continue applying the fluorophore reaction solution until suitable intensity is achieved before moving to the next step.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntibody Elution\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFor paraffin sections, place slides into antigen retrieval solution and incubate in a 95°C water bath for 25–40 min (adjust the time according to antibody affinity), or use mIHC-specific antibody stripping buffer (Cat#C2208) as follows:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply an appropriate amount of prewarmed (37°C) mIHC-specific antibody stripping buffer to cover the sample\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply fresh stripping buffer again\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ewash with PBS 3 times for 5 min each\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eParaffin sections may use either heat retrieval elution or mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eCells (cell climbing slides \/ cell smears), frozen sections, and fragile bone tissue must use the mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecond-round Labeling\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–7.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eDAPI Nuclear Counterstaining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add ready-to-use DAPI staining solution within the marked area and incubate at room temperature for 5–20 min, protected from light.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"6\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMounting\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, mount with the anti-fade mounting medium.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"7\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMicroscope and Imaging\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eObserve and acquire images using:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003efluorescence microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003econfocal microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emultichannel fluorescence scanner\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emultispectral imaging system\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Fluorophore Spectral Information --\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important; color: #000 !important; font-weight: 700 !important;\"\u003e\u003cstrong\u003eFLUOROPHORE SPECTRAL DATA\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; margin: 0px; table-layout: fixed; background: transparent !important; color: rgb(0, 0, 0) !important; height: 163px;\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 31.7827%;\"\u003e \u003ccol style=\"width: 42.7912%;\"\u003e \u003ccol style=\"width: 25.3906%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eProduct Name\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: nowrap; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eExcitation (nm)\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eEmission (nm)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e450\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e520\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-540 Plus\u003cmeta charset=\"utf-8\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e515\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e540\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e550\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e570\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e590\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e620\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-650 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e630\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e650\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e640\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e690\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003eTSA-780 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e750\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e780\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 18.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003eDAPI\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e350\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e420\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003c!-- Precautions --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRECAUTIONS\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e1. Light protection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit have strong anti-photobleaching properties. Full protection from room light is \u003cstrong\u003eNOT\u003c\/strong\u003e required during the workflow, and operation in complete darkness is unnecessary. However, do not expose the reagents or samples to direct sunlight.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e2. Causes of channel crosstalk\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eRelated to the filter bandwidth of the imaging equipment — use narrower bandwidth filters whenever possible.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eThe antibody from the previous round was not completely eluted — optimize elution conditions for high-affinity antibodies, such as increasing elution temperature or time.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSignal imbalance — if two adjacent channels have dyes where one signal is too strong and the other too weak, the stronger signal may bleed into the weaker channel.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e3. Choice of antigen retrieval \/ antibody elution conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eFirst round antigen retrieval: pH 9.0 EDTA, 95°C for 15–25 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSecond and subsequent rounds antigen retrieval \/ antibody elution: pH 6.0 citrate, 95°C for 25–40 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFor tissues that detach easily from the slide, antibody elution buffer may be used instead, but time and temperature must be controlled carefully. Excessive elution time or overly high temperature may reduce antigen recognition or weaken DAPI nuclear staining.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e4. Biomarker \/ antibody order selection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eIn the first round, choose markers suitable for pH 9.0 EDTA retrieval.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eStain the more difficult markers in the earlier rounds.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eAntibodies that are difficult to elute should be used in the last round to avoid crosstalk.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e5. Causes of non-specific staining\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ePolyclonal antibodies are more likely to produce non-specific staining; switch to monoclonal antibodies or reduce concentration and retrieval intensity.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eSignal amplification may be too strong; reduce fluorophore reaction time or concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ePrimary antibody concentration may be too high or retrieval may be excessive; use a higher antibody dilution or lower retrieval strength such as temperature, time, or buffer pH.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Notes --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0;\"\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003ePlease cite this product as \"IHCT003, Bioss Antibodies\". For example: ‘Co-staining of A and B was performed using the Multiplex Fluorescent IHC Staining Kit (IHCT003, Bioss Antibodies), which relies on the Tyramide Signal Amplification (TSA) technology, following the manufacturer’s instructions.’\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e \u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cmeta charset=\"utf-8\"\u003e\u003cem\u003eFor research use only. Not for use in diagnostic or therapeutic procedures.\u003c\/em\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Bioss","offers":[{"title":"100T","offer_id":48159973474603,"sku":"IHCT003","price":1200.0,"currency_code":"USD","in_stock":true},{"title":"20T","offer_id":50861859209515,"sku":"IHCT003-20T","price":390.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/Screenshot2024-03-12at3.53.57PM.png?v=1710273252"},{"product_id":"pv-1024","title":"Goat Anti-Mouse IgG H\u0026L (HRP polymer) Ready-to-Use Kit","description":"\u003ch1 style=\"text-align: center;\" data-mce-fragment=\"1\"\u003eGoat Anti-Mouse IgG H\u0026amp;L (HRP polymer) Ready-to-Use Kit\u003c\/h1\u003e\n\u003cp\u003e\u003cb data-mce-fragment=\"1\"\u003eCat. No.: \u003c\/b\u003ePV-1024\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eApplication: \u003c\/b\u003eImmunohistochemical (IHC) Staining\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eSize: \u003c\/b\u003e6ml \/ 50ml\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eStorage and Stability: \u003c\/b\u003eStable for 12 months at 2-8°C. Protect from light.\u003c\/p\u003e\n\u003ch3\u003e\n\u003cb data-mce-fragment=\"1\"\u003e\u003c\/b\u003e\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ch3\u003eGeneral Information\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ctable width=\"100%\" style=\"height: 96px; width: 97.8221%;\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"height: 19px; width: 69.9443%;\"\u003eComponent\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003eSize\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 39px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 39px;\"\u003eGoat anti-mouse secondary antibody, HRP polymer conjugated\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 39px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eEndogenous peroxidase blocking solution\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eNormal goat serum\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cblockquote\u003e\u003c\/blockquote\u003e\n\u003cp\u003e\u003cstrong\u003ePrinciple\u003c\/strong\u003e\u003cbr\u003eThe detection system facilitates the identification of mouse IgG antibody bond to an antigen in tissue sections. The specific antibody is pinpointed by a secondary antibody that is conjugated with a horseradish peroxidase (HRP) polymer, designed to specifically recognize mouse immunoglobulins. The polymer attached complex is subsequently made visible under a light microscope using HRP-compatible chromogens, such as diaminobenzidine (DAB).\u003c\/p\u003e\n\u003cp\u003e\u003cbr\u003eThe provided HRP polymer conjugated secondary antibody significantly addresses the limitations commonly experienced with traditional immunohistochemistry (IHC) methods, such as poor or inconsistent antigen staining when identifying low-abundance antigens or in situations of suboptimal antibody-antigen binding. By utilizing an HRP polymer conjugate, the sensitivity of detection is dramatically increased, and the process is simplified. Moreover, the HRP polymer-based amplification method reduces the amount of primary antibodies needed and\u003cbr\u003eshortens the secondary antibodies’ incubation period.\u003c\/p\u003e\n\u003ch3\u003e\u003cb data-mce-fragment=\"1\"\u003eProtocol\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePrepare your assay samples and complete antigen retrieval as per your IHC protocol.\u003c\/li\u003e\n\u003cli\u003eDiscard any residual liquid around the tissue section and add 50-100μl of endogenous peroxidase blocking solution, then incubate in a humidity chamber at room temperature (RT) for 15-20 minutes.\u003c\/li\u003e\n\u003cli\u003eWash the slides three times with PBS (PH 7.4) for 3 minutes each.\u003c\/li\u003e\n\u003cli\u003eAdd 50-100μl of normal goat serum to block non-specific binding sites by incubation for 15-20 minutes in a humidity chamber at RT.\u003c\/li\u003e\n\u003cli\u003eDiscard normal goat serum (it’s not necessary to wash) and apply primary antibody for incubation according to the manufacturer’s recommended protocol.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eRemove the remaining liquid around the tissue section and add 50-100μl of Polymer-HRP goat anti-mouse secondary antibody, followed by a 15-20 minute incubation at RT in a humidity chamber.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eApply HRP-compatible chromogens to tissue according to manufacturer’s recommended instructions.\u003c\/li\u003e\n\u003cli\u003eCounterstain and coverslip with a mounting media for microscopy.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb data-mce-fragment=\"1\"\u003eNotes\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePlease thoroughly read this instruction manual before starting the experiment.\u003c\/li\u003e\n\u003cli\u003ePlease note that any uncareful maneuvers within the IHC assay may impact the final results.\u003c\/li\u003e\n\u003cli\u003eNegative results from an IHC staining only indicate that the target antigen was not detected. It would be inappropriate to conclude that the antigen of interest is absent in the samples tested.\u003c\/li\u003e\n\u003cli\u003eBe vigilant to avoid skin and eyes contact with the reagent. Should any contact occur, immediately rinse the\u003cbr\u003eaffected area with plenty of water.\u003c\/li\u003e\n\u003cli\u003eThe HRP-polymer system can be used in IHC autostainers.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003eImportant Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.\u003c\/p\u003e","brand":"Bioss","offers":[{"title":"6ml","offer_id":48371305709867,"sku":"PV-1024-6ml","price":99.0,"currency_code":"USD","in_stock":true},{"title":"50ml","offer_id":48371305742635,"sku":"PV-1024-50ml","price":429.0,"currency_code":"USD","in_stock":true}]},{"product_id":"pv-1023","title":"Goat Anti-Rabbit IgG H\u0026L (HRP polymer) Ready-to-Use Kit","description":"\u003ch1 data-mce-fragment=\"1\" style=\"text-align: center;\"\u003eGoat Anti-Rabbit IgG H\u0026amp;L (HRP polymer) Ready-to-Use Kit\u003c\/h1\u003e\n\u003cp\u003e\u003cb data-mce-fragment=\"1\"\u003eCat. No.: \u003c\/b\u003ePV-1023\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eApplication: \u003c\/b\u003eImmunohistochemical (IHC) Staining\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eSize: \u003c\/b\u003e6ml \/ 50ml\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eStorage and Stability: \u003c\/b\u003eStable for 12 months at 2-8°C. Protect from light.\u003c\/p\u003e\n\u003ch3\u003e\n\u003cb data-mce-fragment=\"1\"\u003e\u003c\/b\u003e\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ch3\u003eGeneral Information\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ctable width=\"100%\" style=\"height: 96px; width: 97.8221%;\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"height: 19px; width: 69.9443%;\"\u003eComponent\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003eSize\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 39px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 39px;\"\u003eGoat anti-rabbit secondary antibody, HRP polymer conjugated\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 39px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eEndogenous peroxidase blocking solution\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eNormal goat serum\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cblockquote\u003e\u003c\/blockquote\u003e\n\u003cp\u003e\u003cstrong\u003ePrinciple\u003c\/strong\u003e\u003cbr\u003eThe detection system facilitates the identification of rabbit IgG antibodies bonded to an antigen in tissue sections. The specific antibody is pinpointed by a secondary antibody that is conjugated with a horseradish peroxidase (HRP) polymer, designed to specifically recognize rabbit immunoglobulins. The polymer-attached complex is subsequently made visible under a light microscope using HRP-compatible chromogens, such as diaminobenzidine (DAB).\u003c\/p\u003e\n\u003cp\u003e\u003cbr data-mce-fragment=\"1\"\u003eThe provided HRP polymer conjugated secondary antibody significantly addresses the limitations commonly experienced with traditional immunohistochemistry (IHC) methods, such as poor or inconsistent antigen staining when identifying low-abundance antigens or in situations of suboptimal antibody-antigen binding. By utilizing an HRP polymer conjugate, the sensitivity of detection is dramatically increased, and the process is simplified. Moreover, the HRP polymer-based amplification method reduces the amount of primary antibodies needed and\u003cbr data-mce-fragment=\"1\"\u003eshortens the secondary antibodies’ incubation period.\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb data-mce-fragment=\"1\"\u003eProtocol\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePrepare your assay samples and complete antigen retrieval as per your IHC protocol.\u003c\/li\u003e\n\u003cli\u003eDiscard any residual liquid around the tissue section and add 50-100μl of endogenous peroxidase blocking solution, then incubate in a humidity chamber at room temperature (RT) for 15-20 minutes.\u003c\/li\u003e\n\u003cli\u003eWash the slides three times with PBS (PH 7.4) for 3 minutes each.\u003c\/li\u003e\n\u003cli\u003eAdd 50-100μl of normal goat serum to block non-specific binding sites by incubation for 15-20 minutes in a humidity chamber at RT.\u003c\/li\u003e\n\u003cli\u003eDiscard normal goat serum (it’s not necessary to wash) and apply primary antibody for incubation according to the manufacturer’s recommended protocol.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eRemove the remaining liquid around the tissue section and add 50-100μl of Polymer-HRP goat anti-mouse secondary antibody, followed by a 15-20 minute incubation at RT in a humidity chamber.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eApply HRP-compatible chromogens to tissue according to manufacturer’s recommended instructions.\u003c\/li\u003e\n\u003cli\u003eCounterstain and coverslip with a mounting media for microscopy.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb data-mce-fragment=\"1\"\u003eNotes\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePlease thoroughly read this instruction manual before starting the experiment.\u003c\/li\u003e\n\u003cli\u003ePlease note that any uncareful maneuvers within the IHC assay may impact the final results.\u003c\/li\u003e\n\u003cli\u003eNegative results from an IHC staining only indicate that the target antigen was not detected. It would be inappropriate to conclude that the antigen of interest is absent in the samples tested.\u003c\/li\u003e\n\u003cli\u003eBe vigilant to avoid skin and eyes contact with the reagent. Should any contact occur, immediately rinse the\u003cbr\u003eaffected area with plenty of water.\u003c\/li\u003e\n\u003cli\u003eThe HRP-polymer system can be used in IHC autostainers.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003eImportant Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.\u003c\/p\u003e","brand":"Bioss","offers":[{"title":"6ml","offer_id":48371535905067,"sku":"PV-1023-6ml","price":99.0,"currency_code":"USD","in_stock":true},{"title":"50ml","offer_id":48371535937835,"sku":"PV-1023-50ml","price":429.0,"currency_code":"USD","in_stock":true}]},{"product_id":"pv-1022","title":"Goat Anti-Mouse \u0026 Rabbit IgG H\u0026L (HRP polymer) Ready-to-Use Kit","description":"\u003ch1 data-mce-fragment=\"1\" style=\"text-align: center;\"\u003eGoat Anti-Mouse \u0026amp; Rabbit IgG H\u0026amp;L (HRP polymer) Ready-to-Use Kit\u003c\/h1\u003e\n\u003cp\u003e\u003cb data-mce-fragment=\"1\"\u003eCat. No.: \u003c\/b\u003ePV-1022\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eApplication: \u003c\/b\u003eImmunohistochemical (IHC) Staining\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eSize: \u003c\/b\u003e6ml \/ 50ml\u003cb data-mce-fragment=\"1\"\u003e\u003cbr\u003eStorage and Stability: \u003c\/b\u003eStable for 12 months at 2-8°C. Protect from light.\u003c\/p\u003e\n\u003ch3\u003e\n\u003cb data-mce-fragment=\"1\"\u003e\u003c\/b\u003e\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ch3\u003eGeneral Information\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ctable width=\"100%\" style=\"height: 96px; width: 97.8221%;\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"height: 19px; width: 69.9443%;\"\u003eComponent\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003eSize\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 39px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 39px;\"\u003eGoat anti-mouse\/rabbit secondary antibody (\u003cstrong\u003euniversal\u003c\/strong\u003e), HRP polymer conjugated\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 39px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eEndogenous peroxidase blocking solution\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eNormal goat serum\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cblockquote\u003e\u003c\/blockquote\u003e\n\u003cp\u003e\u003cstrong\u003ePrinciple\u003c\/strong\u003e\u003cbr\u003eThe detection system facilitates the identification of rabbit IgG antibodies bonded to an antigen in tissue sections. The specific antibody is pinpointed by a \u003cstrong\u003eUNIVERSAL\u003c\/strong\u003e secondary antibody that is conjugated with a horseradish peroxidase (HRP) polymer, designed to specifically recognize both mouse and rabbit immunoglobulins. The polymer-attached complex is subsequently made visible under a light microscope using HRP-compatible chromogens, such as diaminobenzidine (DAB).\u003c\/p\u003e\n\u003cp\u003e\u003cbr data-mce-fragment=\"1\"\u003eThe provided HRP polymer conjugated secondary antibody significantly addresses the limitations commonly experienced with traditional immunohistochemistry (IHC) methods, such as poor or inconsistent antigen staining when identifying low-abundance antigens or in situations of suboptimal antibody-antigen binding. By utilizing an HRP polymer conjugate, the sensitivity of detection is dramatically increased, and the process is simplified. Moreover, the HRP polymer-based amplification method reduces the amount of primary antibody needed and shortens the incubation period of the secondary antibodies.\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb data-mce-fragment=\"1\"\u003eProtocol\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePrepare your assay samples and complete antigen retrieval as per your IHC protocol.\u003c\/li\u003e\n\u003cli\u003eDiscard any residual liquid around the tissue section and add 50-100μl of endogenous peroxidase blocking solution, then incubate in a humidity chamber at room temperature (RT) for 15-20 minutes.\u003c\/li\u003e\n\u003cli\u003eWash the slides three times with PBS (PH 7.4) for 3 minutes each.\u003c\/li\u003e\n\u003cli\u003eAdd 50-100μl of normal goat serum to block non-specific binding sites by incubation for 15-20 minutes in a humidity chamber at RT.\u003c\/li\u003e\n\u003cli\u003eDiscard normal goat serum (it’s not necessary to wash) and apply primary antibody for incubation according to the manufacturer’s recommended protocol.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eRemove the remaining liquid around the tissue section and add 50-100μl of Polymer-HRP goat anti-mouse secondary antibody, followed by a 15-20 minute incubation at RT in a humidity chamber.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eApply HRP-compatible chromogens to tissue according to the manufacturer’s recommended instructions.\u003c\/li\u003e\n\u003cli\u003eCounterstain and coverslip with a mounting media for microscopy.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb data-mce-fragment=\"1\"\u003eNotes\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePlease thoroughly read this instruction manual before starting the experiment.\u003c\/li\u003e\n\u003cli\u003ePlease note that any uncareful maneuvers within the IHC assay may impact the final results.\u003c\/li\u003e\n\u003cli\u003eNegative results from an IHC staining only indicate that the target antigen was not detected. It would be inappropriate to conclude that the antigen of interest is absent in the samples tested.\u003c\/li\u003e\n\u003cli\u003eBe vigilant to avoid skin and eye contact with the reagent. Should any contact occur, immediately rinse the affected area with plenty of water.\u003c\/li\u003e\n\u003cli\u003eThe HRP-polymer system can be used in IHC autostainers.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003eImportant Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.\u003c\/p\u003e","brand":"Bioss","offers":[{"title":"6ml","offer_id":48371536101675,"sku":"PV-1022-6ml","price":99.0,"currency_code":"USD","in_stock":true},{"title":"50ml","offer_id":48371536134443,"sku":"PV-1022-50ml","price":429.0,"currency_code":"USD","in_stock":true}]},{"product_id":"pv-1025","title":"Horse Anti-Mouse \u0026 Rabbit IgG H\u0026L (HRP polymer) Ready-to-Use Kit","description":"\u003ch1 style=\"text-align: center;\"\u003eHorse Anti-Mouse \u0026amp; Rabbit IgG H\u0026amp;L (HRP polymer) Ready-to-Use Kit\u003c\/h1\u003e\n\u003cp\u003e\u003cb\u003eCat. No.: \u003c\/b\u003ePV-1025\u003cb\u003e\u003cbr\u003eApplication: \u003c\/b\u003eImmunohistochemical (IHC) Staining\u003cb\u003e\u003cbr\u003eSize: \u003c\/b\u003e6ml \/ 50ml\u003cb\u003e\u003cbr\u003eStorage and Stability: \u003c\/b\u003eStable for 12 months at 2-8°C. Protect from light.\u003c\/p\u003e\n\u003ch3\u003e\n\u003cb\u003e\u003c\/b\u003e\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ch3\u003eGeneral Information\u003cbr\u003e\n\u003c\/h3\u003e\n\u003ctable style=\"height: 96px; width: 97.8221%;\" width=\"100%\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"height: 19px; width: 69.9443%;\"\u003eComponent\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003eSize\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 39px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 39px;\"\u003eHorse anti-mouse\/rabbit secondary antibody (\u003cstrong\u003euniversal\u003c\/strong\u003e), HRP polymer conjugated\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 39px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eEndogenous peroxidase blocking solution\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 19px;\"\u003e\n\u003ctd style=\"width: 69.9443%; height: 19px;\"\u003eNormal horse serum\u003c\/td\u003e\n\u003ctd style=\"width: 28.5714%; height: 19px;\"\u003e6ml \/ 50ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cblockquote\u003e\u003c\/blockquote\u003e\n\u003cp\u003e\u003cstrong\u003ePrinciple\u003c\/strong\u003e\u003cbr\u003eThe detection system facilitates the identification of rabbit IgG antibodies bonded to an antigen in tissue sections. The specific antibody is pinpointed by a \u003cstrong\u003eUNIVERSAL\u003c\/strong\u003e secondary antibody that is conjugated with a horseradish peroxidase (HRP) polymer, designed to specifically recognize both mouse and rabbit immunoglobulins. The polymer-attached complex is subsequently made visible under a light microscope using HRP-compatible chromogens, such as diaminobenzidine (DAB).\u003c\/p\u003e\n\u003cp\u003e\u003cbr\u003eThe provided HRP polymer conjugated secondary antibody significantly addresses the limitations commonly experienced with traditional immunohistochemistry (IHC) methods, such as poor or inconsistent antigen staining when identifying low-abundance antigens or in situations of suboptimal antibody-antigen binding. By utilizing an HRP polymer conjugate, the sensitivity of detection is dramatically increased, and the process is simplified. Moreover, the HRP polymer-based amplification method reduces the amount of primary antibody needed and shortens the incubation period of the secondary antibodies.\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb\u003eProtocol\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePrepare your assay samples and complete antigen retrieval as per your IHC protocol.\u003c\/li\u003e\n\u003cli\u003eDiscard any residual liquid around the tissue section and add 50-100μl of endogenous peroxidase blocking solution, then incubate in a humidity chamber at room temperature (RT) for 15-20 minutes.\u003c\/li\u003e\n\u003cli\u003eWash the slides three times with PBS (PH 7.4) for 3 minutes each.\u003c\/li\u003e\n\u003cli\u003eAdd 50-100μl of normal horse serum to block non-specific binding sites by incubation for 15-20 minutes in a humidity chamber at RT.\u003c\/li\u003e\n\u003cli\u003eDiscard normal horse serum (it’s not necessary to wash) and apply primary antibody for incubation according to the manufacturer’s recommended protocol.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eRemove the remaining liquid around the tissue section and add 50-100μl of Polymer-HRP horse anti-mouse secondary antibody, followed by a 15-20 minute incubation at RT in a humidity chamber.\u003c\/li\u003e\n\u003cli\u003eWash slides 3 times with PBS (PH 7.4) for 5 minutes each.\u003c\/li\u003e\n\u003cli\u003eApply HRP-compatible chromogens to tissue according to the manufacturer’s recommended instructions.\u003c\/li\u003e\n\u003cli\u003eCounterstain and coverslip with a mounting media for microscopy.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003ch3\u003e\u003cb\u003eNotes\u003cbr\u003e\u003c\/b\u003e\u003c\/h3\u003e\n\u003col\u003e\n\u003cli\u003ePlease thoroughly read this instruction manual before starting the experiment.\u003c\/li\u003e\n\u003cli\u003ePlease note that any uncareful maneuvers within the IHC assay may impact the final results.\u003c\/li\u003e\n\u003cli\u003eNegative IHC staining results only indicate that the target antigen was not detected. It would be inappropriate to conclude that the antigen of interest is absent in the samples tested.\u003c\/li\u003e\n\u003cli\u003eBe vigilant to avoid skin and eye contact with the reagent. Should any contact occur, immediately rinse the affected area with plenty of water.\u003c\/li\u003e\n\u003cli\u003eThe HRP-polymer system can be used in IHC autostainers.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003eImportant Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.\u003c\/p\u003e","brand":"Bioss (Woburn)","offers":[{"title":"6 ml","offer_id":50329520144683,"sku":"PV-1025-6ml","price":99.0,"currency_code":"USD","in_stock":true},{"title":"50 ml","offer_id":50329520177451,"sku":"PV-1025-50ml","price":429.0,"currency_code":"USD","in_stock":true}]},{"product_id":"ihct001","title":"Multiplex Fluorescent (3 colors) IHC Staining Kit","description":"\u003cdiv style=\"line-height: 1.15; font-size: 14px; color: #222;\"\u003e\u003cbr\u003e\u003c\/div\u003e\n\u003cdiv style=\"line-height: 1.15; font-size: 14px; color: #222;\"\u003e\n\u003c!-- Title \/ Overview --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch1 style=\"text-align: center; margin: 0 0 10px 0; line-height: 1.1; background: transparent !important;\"\u003eMultiplex Fluorescent (3 colors) IHC Staining Kit\u003c\/h1\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eCat. No.:\u003c\/strong\u003e IHCT001\u003cbr\u003e\u003cstrong\u003eDetection method:\u003c\/strong\u003e Fluorescent\u003cbr\u003e\u003cstrong\u003eSection type:\u003c\/strong\u003e FFPE \u0026amp; Frozen Tissue, cells\u003cbr\u003e\u003cstrong\u003eSize:\u003c\/strong\u003e 20T | 100T\u003cbr\u003e\u003cstrong\u003eStorage and Stability:\u003c\/strong\u003e Store protected from light at 2–8℃ for 12 months.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c!-- Product Components --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch3 style=\"margin: 0 0 8px 0; line-height: 1.1; background: transparent !important;\"\u003eProduct Components\u003c\/h3\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; text-align: left; table-layout: fixed; line-height: 1.1; margin: 0 0 8px 0; background: transparent !important;\" class=\"product-component\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 28.054%;\"\u003e \u003ccol style=\"width: 42.081%;\"\u003e \u003ccol style=\"width: 14.9148%;\"\u003e \u003ccol style=\"width: 15.0923%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent \u0026lt;p style=;\" rowspan=\"2\"\u003eName\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; background: transparent style=;\" rowspan=\"2\"\u003eComponent\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent \u0026lt;p style=;\" colspan=\"2\"\u003eSpecification\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003e20 T\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003e100 T\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eNuclear dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eDAPI (Ready-to-use)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA Buffer\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e3.2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e16mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eHRP Goat Anti-Rabbit\/Mouse Universal Secondary Antibody\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e3.2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e16mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eAntibody Diluent\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e20mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e3% Hydrogen Peroxide\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e20mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eAnti-fade Mounting Medium\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTSA fluorophore working solution = TSA concentrated fluorophore + TSA buffer.\u003cbr\u003eThe dilution ratio should be optimized according to the specific experiment. The recommended range is 1:50 to 1:400, and 1:200 gives the best results in most cases.\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated at room temperature for 1–3 hours, a dye dilution of 1:50–1:200 is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated overnight at 4°C, a dilution of 1:200–1:400 or higher is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the signal is too strong, reduce the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eIf the signal is too weak, increase the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003c!-- Principle --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRINCIPLE\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTyramide Signal Amplification (TSA) is an enzymatic detection method that uses horseradish peroxidase (HRP) to label target proteins in situ. The principle is based on the peroxidase reaction of tyramide. Under the action of HRP and H₂O₂, the tyramide fluorophore substrate becomes activated. The activated fluorophore then covalently binds to tyrosine and other residues on the target protein, causing substantial fluorophore deposition at the antigen–antibody binding site and thereby amplifying the signal.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter each staining round, the previously bound non-covalent antibodies can be removed by heat-induced retrieval or an antibody elution buffer, while the fluorophore remains stably attached to the protein. This enables the next round of staining. Once all antibody incubations are completed, nuclear staining, mounting, and scanning are performed.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eBecause only one antibody system is present in each staining round, there is no need to worry about antibody cross-reactivity or host-species matching between primary and secondary antibodies. This overcomes the species-origin limitations common in traditional immunofluorescence experiments.\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit may be used individually or in combination and can support single staining, double staining, triple staining, and even more multiplex fluorescence labeling.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c!-- Experimental Procedure --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eEXPERIMENTAL PROCEDURE\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSample Preparation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e1) Paraffin sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace slides sequentially into:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene I, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene II, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol I, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol II, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e95% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e85% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e75% ethanol, 5 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen rinse with distilled water.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e2) Frozen sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFix frozen sections for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e3) Cell climbing slides or cell smears\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFix cell samples for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntigen Retrieval\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace tissue sections into a retrieval container filled with either:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003epH 9.0 EDTA alkaline antigen retrieval solution, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003epH 6.0 citrate retrieval buffer\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePerform antigen retrieval in a microwave oven (other methods such as pressure cooker 1–2 min, boiling water at 100°C for 15 min, or water bath at 95°C for 20 min may also be used).\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eMicrowave program:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emedium heat for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003estop heating and stay still for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emedium-low heat for 7 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eDuring this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter natural cooling, place slides into PBS (pH 7.4), and wash on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe choice of retrieval buffer and conditions depends on tissue type, fixation method, and antigen type. This step may be omitted for frozen sections and cell samples.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Endogenous Peroxidase\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eIncubate the slides in 3% H₂O₂ solution at room temperature, protected from light, for 15 min. Then wash in PBS (pH 7.4) on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Non-specific Binding\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter gently removing excess liquid, draw a hydrophobic barrier around the tissue with a PAP pen.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd antibody diluent (or another blocking reagent such as 3% BSA or goat serum) to completely cover the tissue and block at room temperature for 30 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The antibody diluent contains various stabilizers and preservatives and can be used for blocking or for dilution of primary antibodies. Diluted primary antibodies can be stored long-term at 4°C and for up to one month at room temperature.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003ePrimary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eGently remove the blocking solution and add the appropriately diluted primary antibody working solution to the slide. Incubate flat in a humidified chamber protected from light:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eovernight at 4°C, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e1–2 h at 37°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd a small amount of water into the humid chamber to prevent antibody evaporation.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecondary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add the HRP-conjugated secondary antibody corresponding to the host species of the primary antibody to fully cover the tissue. Incubate at room temperature for 50 min, protected from light, then wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The kit includes a ready-to-use HRP goat anti-rabbit\/mouse universal secondary antibody with very high sensitivity. No preparation is required.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eFluorescence Staining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd the diluted TSA fluorophore working solution to completely cover the tissue and incubate at room temperature for 1–15 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended reaction time: 5–10 min\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e In a pilot experiment, stain for 1 min first, wash, and observe the result. If the positive signal is weak, continue applying the fluorophore reaction solution until suitable intensity is achieved before moving to the next step.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntibody Elution\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFor paraffin sections, place slides into antigen retrieval solution and incubate in a 95°C water bath for 25–40 min (adjust the time according to antibody affinity), or use mIHC-specific antibody stripping buffer (Cat#C2208) as follows:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply an appropriate amount of prewarmed (37°C) mIHC-specific antibody stripping buffer to cover the sample\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply fresh stripping buffer again\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ewash with PBS 3 times for 5 min each\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eParaffin sections may use either heat retrieval elution or mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eCells (cell climbing slides \/ cell smears), frozen sections, and fragile bone tissue must use the mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecond-round Labeling\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–7.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eDAPI Nuclear Counterstaining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add ready-to-use DAPI staining solution within the marked area and incubate at room temperature for 5–20 min, protected from light.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"6\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMounting\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, mount with the anti-fade mounting medium.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"7\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMicroscope and Imaging\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eObserve and acquire images using:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003efluorescence microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003econfocal microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emultichannel fluorescence scanner\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emultispectral imaging system\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003c!-- Fluorophore Spectral Information --\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important; color: #000 !important; font-weight: 700 !important;\"\u003e\u003cstrong\u003eFLUOROPHORE SPECTRAL DATA\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; margin: 0px; table-layout: fixed; background: transparent !important; color: rgb(0, 0, 0) !important; height: 163px;\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 31.7827%;\"\u003e \u003ccol style=\"width: 42.7912%;\"\u003e \u003ccol style=\"width: 25.3906%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eProduct Name\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: nowrap; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eExcitation (nm)\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eEmission (nm)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e450\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e520\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-540 Plus\u003cmeta charset=\"utf-8\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e515\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e540\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e550\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e570\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e590\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e620\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-650 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e630\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e650\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e640\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e690\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003eTSA-780 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e750\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e780\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 18.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003eDAPI\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e350\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e420\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c!-- Precautions --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRECAUTIONS\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e1. Light protection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit have strong anti-photobleaching properties. Full protection from room light is \u003cstrong\u003eNOT\u003c\/strong\u003e required during the workflow, and operation in complete darkness is unnecessary. However, do not expose the reagents or samples to direct sunlight.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e2. Causes of channel crosstalk\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eRelated to the filter bandwidth of the imaging equipment — use narrower bandwidth filters whenever possible.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eThe antibody from the previous round was not completely eluted — optimize elution conditions for high-affinity antibodies, such as increasing elution temperature or time.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSignal imbalance — if two adjacent channels have dyes where one signal is too strong and the other too weak, the stronger signal may bleed into the weaker channel.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e3. Choice of antigen retrieval \/ antibody elution conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eFirst round antigen retrieval: pH 9.0 EDTA, 95°C for 15–25 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSecond and subsequent rounds antigen retrieval \/ antibody elution: pH 6.0 citrate, 95°C for 25–40 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFor tissues that detach easily from the slide, antibody elution buffer may be used instead, but time and temperature must be controlled carefully. Excessive elution time or overly high temperature may reduce antigen recognition or weaken DAPI nuclear staining.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e4. Biomarker \/ antibody order selection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eIn the first round, choose markers suitable for pH 9.0 EDTA retrieval.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eStain the more difficult markers in the earlier rounds.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eAntibodies that are difficult to elute should be used in the last round to avoid crosstalk.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e5. Causes of non-specific staining\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ePolyclonal antibodies are more likely to produce non-specific staining; switch to monoclonal antibodies or reduce concentration and retrieval intensity.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eSignal amplification may be too strong; reduce fluorophore reaction time or concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ePrimary antibody concentration may be too high or retrieval may be excessive; use a higher antibody dilution or lower retrieval strength such as temperature, time, or buffer pH.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003c!-- Notes --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0;\"\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003ePlease cite this product as \"IHCT001, Bioss Antibodies\". For example: ‘Co-staining of A and B was performed using the Multiplex Fluorescent IHC Staining Kit (IHCT001, Bioss Antibodies), which relies on the Tyramide Signal Amplification (TSA) technology, following the manufacturer’s instructions.’\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e \u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cmeta charset=\"utf-8\"\u003e\u003cem\u003eFor research use only. Not for use in diagnostic or therapeutic procedures.\u003c\/em\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Bioss","offers":[{"title":"20T","offer_id":50861830897963,"sku":"IHCT001","price":240.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":50859220533547,"sku":"IHCT001","price":700.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/Screenshot2025-07-01at9.16.34AM.png?v=1751375849"},{"product_id":"ihct002","title":"Multiplex Fluorescent (4 colors) IHC Staining Kit","description":"\u003cdiv style=\"line-height: 1.15; font-size: 14px; color: #222;\"\u003e\n\u003c!-- Title \/ Overview --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch1 style=\"text-align: center; margin: 0 0 10px 0; line-height: 1.1; background: transparent !important;\"\u003eMultiplex Fluorescent (4 colors) IHC Staining Kit\u003c\/h1\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eCat. No.:\u003c\/strong\u003e IHCT002\u003cbr\u003e\u003cstrong\u003eDetection method:\u003c\/strong\u003e Fluorescent\u003cbr\u003e\u003cstrong\u003eSection type:\u003c\/strong\u003e FFPE \u0026amp; Frozen Tissue, cells\u003cbr\u003e\u003cstrong\u003eSize:\u003c\/strong\u003e 20T | 100T\u003cbr\u003e\u003cstrong\u003eStorage and Stability:\u003c\/strong\u003e Store protected from light at 2–8℃ for 12 months.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c!-- Product Components --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch3 style=\"margin: 0 0 8px 0; line-height: 1.1; background: transparent !important;\"\u003eProduct Components\u003c\/h3\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; text-align: left; table-layout: fixed; line-height: 1.1; margin: 0 0 8px 0; background: transparent !important; color: #000 !important;\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 24%;\"\u003e \u003ccol style=\"width: 42%;\"\u003e \u003ccol style=\"width: 17%;\"\u003e \u003ccol style=\"width: 17%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003cth rowspan=\"2\" style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003eName\u003c\/th\u003e\n\u003cth rowspan=\"2\" style=\"border-bottom: 2px solid #333; padding: 6px; background: transparent style=;\"\u003eComponent\u003c\/th\u003e\n\u003cth colspan=\"2\" style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003eSpecification\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003e20T\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid #333; padding: 6px; white-space: nowrap; background: transparent style=;\"\u003e100T\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10μL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e50μL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10μL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e50μL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10μL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e50μL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eNuclear dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eDAPI (Ready-to-use)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA Buffer\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e4.8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e24mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eHRP Goat Anti-Rabbit\/Mouse Universal Secondary Antibody\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e4.8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e24mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eAntibody Diluent\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e6mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e30mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e3% Hydrogen Peroxide\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e6mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e30mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eReagent 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eAnti-fade Mounting Medium\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTSA fluorophore working solution = TSA concentrated fluorophore + TSA buffer.\u003cbr\u003eThe dilution ratio should be optimized according to the specific experiment. The recommended range is 1:50 to 1:400, and 1:200 gives the best results in most cases.\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated at room temperature for 1–3 hours, a dye dilution of 1:50–1:200 is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated overnight at 4°C, a dilution of 1:200–1:400 or higher is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the signal is too strong, reduce the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eIf the signal is too weak, increase the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003c!-- Principle --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRINCIPLE\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTyramide Signal Amplification (TSA) is an enzymatic detection method that uses horseradish peroxidase (HRP) to label target proteins in situ. The principle is based on the peroxidase reaction of tyramide. Under the action of HRP and H₂O₂, the tyramide fluorophore substrate becomes activated. The activated fluorophore then covalently binds to tyrosine and other residues on the target protein, causing substantial fluorophore deposition at the antigen–antibody binding site and thereby amplifying the signal.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter each staining round, the previously bound non-covalent antibodies can be removed by heat-induced retrieval or an antibody elution buffer, while the fluorophore remains stably attached to the protein. This enables the next round of staining. Once all antibody incubations are completed, nuclear staining, mounting, and scanning are performed.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eBecause only one antibody system is present in each staining round, there is no need to worry about antibody cross-reactivity or host-species matching between primary and secondary antibodies. This overcomes the species-origin limitations common in traditional immunofluorescence experiments.\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit may be used individually or in combination and can support single staining, double staining, triple staining, and even more multiplex fluorescence labeling.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c!-- Experimental Procedure --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eEXPERIMENTAL PROCEDURE\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSample Preparation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e1) Paraffin sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace slides sequentially into:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene I, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene II, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol I, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol II, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e95% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e85% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e75% ethanol, 5 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen rinse with distilled water.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e2) Frozen sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFix frozen sections for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e3) Cell climbing slides or cell smears\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFix cell samples for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003col start=\"2\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntigen Retrieval\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace tissue sections into a retrieval container filled with either:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003epH 9.0 EDTA alkaline antigen retrieval solution, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003epH 6.0 citrate retrieval buffer\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePerform antigen retrieval in a microwave oven (other methods such as pressure cooker 1–2 min, boiling water at 100°C for 15 min, or water bath at 95°C for 20 min may also be used).\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eMicrowave program:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emedium heat for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003estop heating and stay still for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emedium-low heat for 7 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eDuring this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter natural cooling, place slides into PBS (pH 7.4), and wash on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe choice of retrieval buffer and conditions depends on tissue type, fixation method, and antigen type. This step may be omitted for frozen sections and cell samples.\u003c\/p\u003e\n\u003col start=\"3\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Endogenous Peroxidase\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eIncubate the slides in 3% H₂O₂ solution at room temperature, protected from light, for 15 min. Then wash in PBS (pH 7.4) on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003col start=\"4\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Non-specific Binding\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter gently removing excess liquid, draw a hydrophobic barrier around the tissue with a PAP pen.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd antibody diluent (or another blocking reagent such as 3% BSA or goat serum) to completely cover the tissue and block at room temperature for 30 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The antibody diluent contains various stabilizers and preservatives and can be used for blocking or for dilution of primary antibodies. Diluted primary antibodies can be stored long-term at 4°C and for up to one month at room temperature.\u003c\/p\u003e\n\u003col start=\"5\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003ePrimary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eGently remove the blocking solution and add the appropriately diluted primary antibody working solution to the slide. Incubate flat in a humidified chamber protected from light:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eovernight at 4°C, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e1–2 h at 37°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd a small amount of water into the humid chamber to prevent antibody evaporation.\u003c\/p\u003e\n\u003col start=\"6\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecondary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add the HRP-conjugated secondary antibody corresponding to the host species of the primary antibody to fully cover the tissue. Incubate at room temperature for 50 min, protected from light, then wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The kit includes a ready-to-use HRP goat anti-rabbit\/mouse universal secondary antibody with very high sensitivity. No preparation is required.\u003c\/p\u003e\n\u003col start=\"7\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eFluorescence Staining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd the diluted TSA fluorophore working solution to completely cover the tissue and incubate at room temperature for 1–15 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended reaction time: 5–10 min\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e In a pilot experiment, stain for 1 min first, wash, and observe the result. If the positive signal is weak, continue applying the fluorophore reaction solution until suitable intensity is achieved before moving to the next step.\u003c\/p\u003e\n\u003col start=\"8\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntibody Elution\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFor paraffin sections, place slides into antigen retrieval solution and incubate in a 95°C water bath for 25–40 min (adjust the time according to antibody affinity), or use mIHC-specific antibody stripping buffer (Cat#C2208) as follows:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply an appropriate amount of prewarmed (37°C) mIHC-specific antibody stripping buffer to cover the sample\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply fresh stripping buffer again\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ewash with PBS 3 times for 5 min each\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eParaffin sections may use either heat retrieval elution or mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eCells (cell climbing slides \/ cell smears), frozen sections, and fragile bone tissue must use the mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003col start=\"9\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecond-round Labeling\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–8.\u003c\/p\u003e\n\u003col start=\"10\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eThird-round Labeling\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–7.\u003c\/p\u003e\n\u003col start=\"11\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eDAPI Nuclear Counterstaining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add ready-to-use DAPI staining solution within the marked area and incubate at room temperature for 5–20 min, protected from light.\u003c\/p\u003e\n\u003col start=\"12\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMounting\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, mount with the anti-fade mounting medium.\u003c\/p\u003e\n\u003col start=\"13\" style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMicroscope and Imaging\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eObserve and acquire images using:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003efluorescence microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003econfocal microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emultichannel fluorescence scanner\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emultispectral imaging system\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003c!-- Fluorophore Spectral Information --\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important; color: #000 !important; font-weight: 700 !important;\"\u003e\u003cstrong\u003eFLUOROPHORE SPECTRAL DATA\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; margin: 0; table-layout: fixed; background: transparent !important; color: #000 !important;\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 42%;\"\u003e \u003ccol style=\"width: 29%;\"\u003e \u003ccol style=\"width: 29%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: #9a9a9a !important;\"\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; color: #fff !important; font-weight: bold; background: #9a9a9a !important; border: none;\"\u003eProduct Name\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; color: #fff !important; font-weight: bold; background: #9a9a9a !important; border: none; white-space: nowrap;\"\u003eExcitation (nm)\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; color: #fff !important; font-weight: bold; background: #9a9a9a !important; border: none; white-space: nowrap;\"\u003eEmission (nm)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: #efeff4 !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important;\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e450\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e480\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #ffffff !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important;\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e480\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e520\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #efeff4 !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important;\"\u003eTSA-540 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e515\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e540\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #ffffff !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important;\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e550\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e570\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #efeff4 !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important;\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e590\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e620\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #ffffff !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important;\"\u003eTSA-650 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e630\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e650\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #efeff4 !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important;\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e640\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e690\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #ffffff !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important;\"\u003eTSA-780 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e750\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #ffffff !important; color: #000 !important; white-space: nowrap;\"\u003e780\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: #efeff4 !important;\"\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important;\"\u003eDAPI\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e350\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; background: #efeff4 !important; color: #000 !important; white-space: nowrap;\"\u003e420\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c!-- Precautions --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 10px 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRECAUTIONS\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e1. Light protection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit have strong anti-photobleaching properties. Full protection from room light is \u003cstrong\u003eNOT\u003c\/strong\u003e required during the workflow, and operation in complete darkness is unnecessary. However, do not expose the reagents or samples to direct sunlight.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e2. Causes of channel crosstalk\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eRelated to the filter bandwidth of the imaging equipment — use narrower bandwidth filters whenever possible.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eThe antibody from the previous round was not completely eluted — optimize elution conditions for high-affinity antibodies, such as increasing elution temperature or time.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSignal imbalance — if two adjacent channels have dyes where one signal is too strong and the other too weak, the stronger signal may bleed into the weaker channel.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e3. Choice of antigen retrieval \/ antibody elution conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eFirst round antigen retrieval: pH 9.0 EDTA, 95°C for 15–25 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSecond and subsequent rounds antigen retrieval \/ antibody elution: pH 6.0 citrate, 95°C for 25–40 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFor tissues that detach easily from the slide, antibody elution buffer may be used instead, but time and temperature must be controlled carefully. Excessive elution time or overly high temperature may reduce antigen recognition or weaken DAPI nuclear staining.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e4. Biomarker \/ antibody order selection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eIn the first round, choose markers suitable for pH 9.0 EDTA retrieval.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eStain the more difficult markers in the earlier rounds.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eAntibodies that are difficult to elute should be used in the last round to avoid crosstalk.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e5. Causes of non-specific staining\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ePolyclonal antibodies are more likely to produce non-specific staining; switch to monoclonal antibodies or reduce concentration and retrieval intensity.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eSignal amplification may be too strong; reduce fluorophore reaction time or concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ePrimary antibody concentration may be too high or retrieval may be excessive; use a higher antibody dilution or lower retrieval strength such as temperature, time, or buffer pH.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003c!-- Notes --\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0;\"\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003ePlease cite this product as \"IHCT002, Bioss Antibodies\". For example: ‘Co-staining of A, B and C was performed using the Multiplex Fluorescent IHC Staining Kit (IHCT002, Bioss Antibodies), which relies on the Tyramide Signal Amplification (TSA) technology, following the manufacturer’s instructions.’\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cem\u003eFor research use only. Not for use in diagnostic or therapeutic procedures.\u003c\/em\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e","brand":"Bioss","offers":[{"title":"20T","offer_id":50861832700203,"sku":"IHCT002","price":280.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":50859220697387,"sku":"IHCT002","price":860.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/Screenshot2025-07-01at9.18.11AM.png?v=1751375907"},{"product_id":"ihct004","title":"Multiplex Fluorescent (6 colors) IHC Staining Kit","description":"\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch1 style=\"text-align: center; margin: 0 0 10px 0; line-height: 1.1; background: transparent !important;\"\u003eMultiplex Fluorescent (6 colors) IHC Staining Kit\u003c\/h1\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eCat. No.:\u003c\/strong\u003e IHCT004\u003cbr\u003e\u003cstrong\u003eDetection method:\u003c\/strong\u003e Fluorescent\u003cbr\u003e\u003cstrong\u003eSection type:\u003c\/strong\u003e FFPE \u0026amp; Frozen Tissue, cells\u003cbr\u003e\u003cstrong\u003eSize:\u003c\/strong\u003e 20T | 100T\u003cbr\u003e\u003cstrong\u003eStorage and Stability:\u003c\/strong\u003e Store protected from light at 2–8℃ for 12 months.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Product Components --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch3 style=\"margin: 0 0 8px 0; line-height: 1.1; background: transparent !important;\"\u003eProduct Components\u003c\/h3\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; text-align: left; table-layout: fixed; line-height: 1.1; margin: 0px 0px 8px; background: transparent !important; height: 277.2px;\" class=\"product-component\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 28.0497%;\"\u003e \u003ccol style=\"width: 41.9804%;\"\u003e \u003ccol style=\"width: 14.872%;\"\u003e \u003ccol style=\"width: 15.0602%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 30.8px;\" rowspan=\"2\"\u003eName\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; height: 30.8px;\" rowspan=\"2\"\u003eComponent\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 15.4px;\" colspan=\"2\"\u003eSpecification\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 15.4px;\"\u003e20 T\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 15.4px;\"\u003e100 T\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eTSA dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA dye 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e \u003cbr\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA dye 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA dye 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA dye 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eNuclear dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eDAPI (Ready-to-use)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eTSA Buffer\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e6.4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e32mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eReagent 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eHRP Goat Anti-Rabbit\/Mouse Universal Secondary Antibody\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e6.4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e32mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eAntibody Diluent\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e3% Hydrogen Peroxide\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eAnti-fade Mounting Medium\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTSA fluorophore working solution = TSA concentrated fluorophore + TSA buffer.\u003cbr\u003eThe dilution ratio should be optimized according to the specific experiment. The recommended range is 1:50 to 1:400, and 1:200 gives the best results in most cases.\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated at room temperature for 1–3 hours, a dye dilution of 1:50–1:200 is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated overnight at 4°C, a dilution of 1:200–1:400 or higher is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the signal is too strong, reduce the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eIf the signal is too weak, increase the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Principle --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRINCIPLE\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTyramide Signal Amplification (TSA) is an enzymatic detection method that uses horseradish peroxidase (HRP) to label target proteins in situ. The principle is based on the peroxidase reaction of tyramide. Under the action of HRP and H₂O₂, the tyramide fluorophore substrate becomes activated. The activated fluorophore then covalently binds to tyrosine and other residues on the target protein, causing substantial fluorophore deposition at the antigen–antibody binding site and thereby amplifying the signal.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter each staining round, the previously bound non-covalent antibodies can be removed by heat-induced retrieval or an antibody elution buffer, while the fluorophore remains stably attached to the protein. This enables the next round of staining. Once all antibody incubations are completed, nuclear staining, mounting, and scanning are performed.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eBecause only one antibody system is present in each staining round, there is no need to worry about antibody cross-reactivity or host-species matching between primary and secondary antibodies. This overcomes the species-origin limitations common in traditional immunofluorescence experiments.\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit may be used individually or in combination and can support single staining, double staining, triple staining, and even more multiplex fluorescence labeling.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Experimental Procedure --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eEXPERIMENTAL PROCEDURE\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSample Preparation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e1) Paraffin sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace slides sequentially into:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene I, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene II, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol I, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol II, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e95% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e85% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e75% ethanol, 5 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen rinse with distilled water.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e2) Frozen sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFix frozen sections for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e3) Cell climbing slides or cell smears\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFix cell samples for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntigen Retrieval\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace tissue sections into a retrieval container filled with either:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003epH 9.0 EDTA alkaline antigen retrieval solution, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003epH 6.0 citrate retrieval buffer\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePerform antigen retrieval in a microwave oven (other methods such as pressure cooker 1–2 min, boiling water at 100°C for 15 min, or water bath at 95°C for 20 min may also be used).\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eMicrowave program:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emedium heat for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003estop heating and stay still for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emedium-low heat for 7 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eDuring this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter natural cooling, place slides into PBS (pH 7.4), and wash on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe choice of retrieval buffer and conditions depends on tissue type, fixation method, and antigen type. This step may be omitted for frozen sections and cell samples.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Endogenous Peroxidase\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eIncubate the slides in 3% H₂O₂ solution at room temperature, protected from light, for 15 min. Then wash in PBS (pH 7.4) on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Non-specific Binding\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter gently removing excess liquid, draw a hydrophobic barrier around the tissue with a PAP pen.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd antibody diluent (or another blocking reagent such as 3% BSA or goat serum) to completely cover the tissue and block at room temperature for 30 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The antibody diluent contains various stabilizers and preservatives and can be used for blocking or for dilution of primary antibodies. Diluted primary antibodies can be stored long-term at 4°C and for up to one month at room temperature.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003ePrimary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eGently remove the blocking solution and add the appropriately diluted primary antibody working solution to the slide. Incubate flat in a humidified chamber protected from light:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eovernight at 4°C, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e1–2 h at 37°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd a small amount of water into the humid chamber to prevent antibody evaporation.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecondary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add the HRP-conjugated secondary antibody corresponding to the host species of the primary antibody to fully cover the tissue. Incubate at room temperature for 50 min, protected from light, then wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The kit includes a ready-to-use HRP goat anti-rabbit\/mouse universal secondary antibody with very high sensitivity. No preparation is required.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eFluorescence Staining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd the diluted TSA fluorophore working solution to completely cover the tissue and incubate at room temperature for 1–15 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended reaction time: 5–10 min\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e In a pilot experiment, stain for 1 min first, wash, and observe the result. If the positive signal is weak, continue applying the fluorophore reaction solution until suitable intensity is achieved before moving to the next step.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntibody Elution\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFor paraffin sections, place slides into antigen retrieval solution and incubate in a 95°C water bath for 25–40 min (adjust the time according to antibody affinity), or use mIHC-specific antibody stripping buffer (Cat#C2208) as follows:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply an appropriate amount of prewarmed (37°C) mIHC-specific antibody stripping buffer to cover the sample\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply fresh stripping buffer again\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ewash with PBS 3 times for 5 min each\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eParaffin sections may use either heat retrieval elution or mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eCells (cell climbing slides \/ cell smears), frozen sections, and fragile bone tissue must use the mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecond-round Labeling\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–7.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eDAPI Nuclear Counterstaining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add ready-to-use DAPI staining solution within the marked area and incubate at room temperature for 5–20 min, protected from light.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"6\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMounting\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, mount with the anti-fade mounting medium.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"7\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMicroscope and Imaging\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eObserve and acquire images using:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003efluorescence microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003econfocal microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emultichannel fluorescence scanner\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emultispectral imaging system\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Fluorophore Spectral Information --\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important; color: #000 !important; font-weight: 700 !important;\"\u003e\u003cstrong\u003eFLUOROPHORE SPECTRAL DATA\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; margin: 0px; table-layout: fixed; background: transparent !important; color: rgb(0, 0, 0) !important; height: 163px;\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 31.7827%;\"\u003e \u003ccol style=\"width: 42.7912%;\"\u003e \u003ccol style=\"width: 25.3906%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eProduct Name\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: nowrap; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eExcitation (nm)\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eEmission (nm)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e450\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e520\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-540 Plus\u003cmeta charset=\"utf-8\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e515\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e540\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e550\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e570\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e590\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e620\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-650 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e630\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e650\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e640\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e690\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003eTSA-780 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e750\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e780\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 18.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003eDAPI\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e350\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e420\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003c!-- Precautions --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRECAUTIONS\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e1. Light protection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit have strong anti-photobleaching properties. Full protection from room light is \u003cstrong\u003eNOT\u003c\/strong\u003e required during the workflow, and operation in complete darkness is unnecessary. However, do not expose the reagents or samples to direct sunlight.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e2. Causes of channel crosstalk\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eRelated to the filter bandwidth of the imaging equipment — use narrower bandwidth filters whenever possible.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eThe antibody from the previous round was not completely eluted — optimize elution conditions for high-affinity antibodies, such as increasing elution temperature or time.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSignal imbalance — if two adjacent channels have dyes where one signal is too strong and the other too weak, the stronger signal may bleed into the weaker channel.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e3. Choice of antigen retrieval \/ antibody elution conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eFirst round antigen retrieval: pH 9.0 EDTA, 95°C for 15–25 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSecond and subsequent rounds antigen retrieval \/ antibody elution: pH 6.0 citrate, 95°C for 25–40 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFor tissues that detach easily from the slide, antibody elution buffer may be used instead, but time and temperature must be controlled carefully. Excessive elution time or overly high temperature may reduce antigen recognition or weaken DAPI nuclear staining.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e4. Biomarker \/ antibody order selection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eIn the first round, choose markers suitable for pH 9.0 EDTA retrieval.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eStain the more difficult markers in the earlier rounds.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eAntibodies that are difficult to elute should be used in the last round to avoid crosstalk.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e5. Causes of non-specific staining\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ePolyclonal antibodies are more likely to produce non-specific staining; switch to monoclonal antibodies or reduce concentration and retrieval intensity.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eSignal amplification may be too strong; reduce fluorophore reaction time or concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ePrimary antibody concentration may be too high or retrieval may be excessive; use a higher antibody dilution or lower retrieval strength such as temperature, time, or buffer pH.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Notes --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0;\"\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003ePlease cite this product as \"IHCT004, Bioss Antibodies\". For example: ‘Co-staining of A and B was performed using the Multiplex Fluorescent IHC Staining Kit (IHCT004, Bioss Antibodies), which relies on the Tyramide Signal Amplification (TSA) technology, following the manufacturer’s instructions.’\u003cem\u003e\u003c\/em\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cem\u003eFor research use only. Not for use in diagnostic or therapeutic procedures.\u003c\/em\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Bioss","offers":[{"title":"20T","offer_id":50861833322795,"sku":"IHCT004","price":520.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":50859221188907,"sku":"IHCT004","price":1600.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/Screenshot2025-07-01at9.18.52AM.png?v=1751375947"},{"product_id":"ihct005","title":"Multiplex Fluorescent (7 colors) IHC Staining Kit","description":"\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch1 style=\"text-align: center; margin: 0 0 10px 0; line-height: 1.1; background: transparent !important;\"\u003eMultiplex Fluorescent (7 colors) IHC Staining Kit\u003c\/h1\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eCat. No.:\u003c\/strong\u003e IHCT005\u003cbr\u003e\u003cstrong\u003eDetection method:\u003c\/strong\u003e Fluorescent\u003cbr\u003e\u003cstrong\u003eSection type:\u003c\/strong\u003e FFPE \u0026amp; Frozen Tissue, cells\u003cbr\u003e\u003cstrong\u003eSize:\u003c\/strong\u003e 20T | 100T\u003cbr\u003e\u003cstrong\u003eStorage and Stability:\u003c\/strong\u003e Store protected from light at 2–8℃ for 12 months.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Product Components --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003ch3 style=\"margin: 0 0 8px 0; line-height: 1.1; background: transparent !important;\"\u003eProduct Components\u003c\/h3\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; text-align: left; table-layout: fixed; line-height: 1.1; margin: 0px 0px 8px; background: transparent !important; height: 277.2px;\" class=\"product-component\"\u003e\n\u003ccolgroup\u003e\n\u003ccol style=\"width: 28.0497%;\"\u003e\n\u003ccol style=\"width: 41.9804%;\"\u003e\n\u003ccol style=\"width: 14.872%;\"\u003e\n\u003ccol style=\"width: 15.0602%;\"\u003e\n\u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 30.8px;\" rowspan=\"2\"\u003eName\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; height: 30.8px;\" rowspan=\"2\"\u003eComponent\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 15.4px;\" colspan=\"2\"\u003eSpecification\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 15.4px;\"\u003e20 T\u003c\/th\u003e\n\u003cth style=\"border-bottom: 2px solid rgb(51, 51, 51); padding: 6px; white-space: nowrap; height: 15.4px;\"\u003e100 T\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eTSA dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA dye 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e \u003cbr\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA dye 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA dye 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA dye 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA dye 6 \u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003eTSA-780 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e10µL (200X)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003e50µL (200X)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eNuclear dye 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eDAPI (Ready-to-use)\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 1\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eTSA Buffer\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e6.4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e32mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 30.8px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eReagent 2\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003eHRP Goat Anti-Rabbit\/Mouse Universal Secondary Antibody\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 30.8px;\"\u003e6.4mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 30.8px;\"\u003e32mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 3\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eAntibody Diluent\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 4\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e3% Hydrogen Peroxide\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e8mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e40mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: transparent !important; height: 15.4px;\"\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eReagent 5\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003eAnti-fade Mounting Medium\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; text-align: center; background: transparent !important; height: 15.4px;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"padding: 6px; background: transparent !important; height: 15.4px;\"\u003e10mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTSA fluorophore working solution = TSA concentrated fluorophore + TSA buffer.\u003cbr\u003eThe dilution ratio should be optimized according to the specific experiment. The recommended range is 1:50 to 1:400, and 1:200 gives the best results in most cases.\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated at room temperature for 1–3 hours, a dye dilution of 1:50–1:200 is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the primary antibody is incubated overnight at 4°C, a dilution of 1:200–1:400 or higher is recommended.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eIf the signal is too strong, reduce the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eIf the signal is too weak, increase the dye concentration, reaction time, or primary antibody concentration.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Principle --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRINCIPLE\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eTyramide Signal Amplification (TSA) is an enzymatic detection method that uses horseradish peroxidase (HRP) to label target proteins in situ. The principle is based on the peroxidase reaction of tyramide. Under the action of HRP and H₂O₂, the tyramide fluorophore substrate becomes activated. The activated fluorophore then covalently binds to tyrosine and other residues on the target protein, causing substantial fluorophore deposition at the antigen–antibody binding site and thereby amplifying the signal.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter each staining round, the previously bound non-covalent antibodies can be removed by heat-induced retrieval or an antibody elution buffer, while the fluorophore remains stably attached to the protein. This enables the next round of staining. Once all antibody incubations are completed, nuclear staining, mounting, and scanning are performed.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eBecause only one antibody system is present in each staining round, there is no need to worry about antibody cross-reactivity or host-species matching between primary and secondary antibodies. This overcomes the species-origin limitations common in traditional immunofluorescence experiments.\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit may be used individually or in combination and can support single staining, double staining, triple staining, and even more multiplex fluorescence labeling.\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Experimental Procedure --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eEXPERIMENTAL PROCEDURE\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSample Preparation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e1) Paraffin sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace slides sequentially into:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene I, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene II, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol I, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol II, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e95% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e85% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e75% ethanol, 5 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen rinse with distilled water.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e2) Frozen sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFix frozen sections for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e3) Cell climbing slides or cell smears\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFix cell samples for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntigen Retrieval\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace tissue sections into a retrieval container filled with either:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003epH 9.0 EDTA alkaline antigen retrieval solution, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003epH 6.0 citrate retrieval buffer\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePerform antigen retrieval in a microwave oven (other methods such as pressure cooker 1–2 min, boiling water at 100°C for 15 min, or water bath at 95°C for 20 min may also be used).\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eMicrowave program:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emedium heat for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003estop heating and stay still for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emedium-low heat for 7 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eDuring this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter natural cooling, place slides into PBS (pH 7.4), and wash on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe choice of retrieval buffer and conditions depends on tissue type, fixation method, and antigen type. This step may be omitted for frozen sections and cell samples.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Endogenous Peroxidase\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eIncubate the slides in 3% H₂O₂ solution at room temperature, protected from light, for 15 min. Then wash in PBS (pH 7.4) on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Non-specific Binding\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter gently removing excess liquid, draw a hydrophobic barrier around the tissue with a PAP pen.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd antibody diluent (or another blocking reagent such as 3% BSA or goat serum) to completely cover the tissue and block at room temperature for 30 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The antibody diluent contains various stabilizers and preservatives and can be used for blocking or for dilution of primary antibodies. Diluted primary antibodies can be stored long-term at 4°C and for up to one month at room temperature.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003ePrimary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eGently remove the blocking solution and add the appropriately diluted primary antibody working solution to the slide. Incubate flat in a humidified chamber protected from light:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eovernight at 4°C, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e1–2 h at 37°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd a small amount of water into the humid chamber to prevent antibody evaporation.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecondary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add the HRP-conjugated secondary antibody corresponding to the host species of the primary antibody to fully cover the tissue. Incubate at room temperature for 50 min, protected from light, then wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The kit includes a ready-to-use HRP goat anti-rabbit\/mouse universal secondary antibody with very high sensitivity. No preparation is required.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eFluorescence Staining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd the diluted TSA fluorophore working solution to completely cover the tissue and incubate at room temperature for 1–15 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended reaction time: 5–10 min\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e In a pilot experiment, stain for 1 min first, wash, and observe the result. If the positive signal is weak, continue applying the fluorophore reaction solution until suitable intensity is achieved before moving to the next step.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntibody Elution\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFor paraffin sections, place slides into antigen retrieval solution and incubate in a 95°C water bath for 25–40 min (adjust the time according to antibody affinity), or use mIHC-specific antibody stripping buffer (Cat#C2208) as follows:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply an appropriate amount of prewarmed (37°C) mIHC-specific antibody stripping buffer to cover the sample\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eapply fresh stripping buffer again\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eincubate at 37°C for 5–20 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ediscard the stripping buffer\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ewash with PBS 3 times for 5 min each\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eParaffin sections may use either heat retrieval elution or mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eCells (cell climbing slides \/ cell smears), frozen sections, and fragile bone tissue must use the mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSecond-round Labeling\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–7.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eDAPI Nuclear Counterstaining\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add ready-to-use DAPI staining solution within the marked area and incubate at room temperature for 5–20 min, protected from light.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"6\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMounting\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, mount with the anti-fade mounting medium.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"7\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eMicroscope and Imaging\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eObserve and acquire images using:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003efluorescence microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003econfocal microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emultichannel fluorescence scanner\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emultispectral imaging system\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Fluorophore Spectral Information --\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important; color: #000 !important; font-weight: 700 !important;\"\u003e\u003cstrong\u003eFLUOROPHORE SPECTRAL DATA\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"width: 100%; border-collapse: collapse; margin: 0px; table-layout: fixed; background: transparent !important; color: rgb(0, 0, 0) !important; height: 163px;\"\u003e\n\u003ccolgroup\u003e \u003ccol style=\"width: 31.7827%;\"\u003e \u003ccol style=\"width: 42.7912%;\"\u003e \u003ccol style=\"width: 25.3906%;\"\u003e \u003c\/colgroup\u003e\n\u003cthead style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eProduct Name\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: nowrap; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eExcitation (nm)\u003c\/th\u003e\n\u003cth style=\"padding: 10px 12px; text-align: left; font-weight: bold; border: none; white-space: normal; vertical-align: middle; color: rgb(255, 255, 255) !important; background: rgb(154, 154, 154) !important; height: 16.1px;\"\u003eEmission (nm)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003c\/thead\u003e\n\u003ctbody style=\"background: transparent !important;\"\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-480 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e450\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-520 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e480\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e520\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-540 Plus\u003cmeta charset=\"utf-8\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e515\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e540\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-570 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e550\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e570\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-620 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e590\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e620\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-650 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e630\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e650\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e\n\u003cmeta charset=\"utf-8\"\u003eTSA-690 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e640\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e690\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(255, 255, 255) !important; height: 16.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003eTSA-780 Plus\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e750\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(255, 255, 255) !important; color: rgb(0, 0, 0) !important; height: 16.1px;\"\u003e780\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"background: rgb(239, 239, 244) !important; height: 18.1px;\"\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003eDAPI\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e350\u003c\/td\u003e\n\u003ctd style=\"padding: 12px; vertical-align: top; white-space: nowrap; background: rgb(239, 239, 244) !important; color: rgb(0, 0, 0) !important; height: 18.1px;\"\u003e420\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003c!-- Precautions --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRECAUTIONS\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e1. Light protection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe fluorophores in this kit have strong anti-photobleaching properties. Full protection from room light is \u003cstrong\u003eNOT\u003c\/strong\u003e required during the workflow, and operation in complete darkness is unnecessary. However, do not expose the reagents or samples to direct sunlight.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e2. Causes of channel crosstalk\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eRelated to the filter bandwidth of the imaging equipment — use narrower bandwidth filters whenever possible.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eThe antibody from the previous round was not completely eluted — optimize elution conditions for high-affinity antibodies, such as increasing elution temperature or time.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSignal imbalance — if two adjacent channels have dyes where one signal is too strong and the other too weak, the stronger signal may bleed into the weaker channel.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e3. Choice of antigen retrieval \/ antibody elution conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eFirst round antigen retrieval: pH 9.0 EDTA, 95°C for 15–25 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eSecond and subsequent rounds antigen retrieval \/ antibody elution: pH 6.0 citrate, 95°C for 25–40 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFor tissues that detach easily from the slide, antibody elution buffer may be used instead, but time and temperature must be controlled carefully. Excessive elution time or overly high temperature may reduce antigen recognition or weaken DAPI nuclear staining.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e4. Biomarker \/ antibody order selection\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eIn the first round, choose markers suitable for pH 9.0 EDTA retrieval.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eStain the more difficult markers in the earlier rounds.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eAntibodies that are difficult to elute should be used in the last round to avoid crosstalk.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e5. Causes of non-specific staining\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003ePolyclonal antibodies are more likely to produce non-specific staining; switch to monoclonal antibodies or reduce concentration and retrieval intensity.\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eSignal amplification may be too strong; reduce fluorophore reaction time or concentration.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003ePrimary antibody concentration may be too high or retrieval may be excessive; use a higher antibody dilution or lower retrieval strength such as temperature, time, or buffer pH.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Notes --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0;\"\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003ePlease cite this product as \"IHCT005, Bioss Antibodies\". For example: ‘Co-staining of A and B was performed using the Multiplex Fluorescent IHC Staining Kit (IHCT005, Bioss Antibodies), which relies on the Tyramide Signal Amplification (TSA) technology, following the manufacturer’s instructions.’\u003cem\u003e\u003c\/em\u003e\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cem\u003eFor research use only. Not for use in diagnostic or therapeutic procedures.\u003c\/em\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Bioss","offers":[{"title":"20T","offer_id":50861833453867,"sku":"IHCT005","price":720.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":50859221516587,"sku":"IHCT005","price":2300.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/Screenshot2025-07-01at9.19.35AM.png?v=1751376021"},{"product_id":"c2208","title":"mIHC-specific Antibody Stripping Buffer","description":"\u003cdiv\u003e\n\u003cbr\u003e\n\u003ch1 style=\"text-align: center; margin: 0 0 10px 0; line-height: 1.1; background: transparent !important;\"\u003emIHC-specific Antibody Stripping Buffer\u003c\/h1\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eCat. No.:\u003c\/strong\u003e C2208\u003cbr\u003e\u003cstrong\u003eSize:\u003c\/strong\u003e 30mL\u003cbr\u003e\u003cstrong\u003eStorage and Stability:\u003c\/strong\u003e Store at room temperature for 12 months.\u003c\/p\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cmeta charset=\"utf-8\"\u003e\u003cb\u003eApplication:\u003c\/b\u003e Suitable for paraffin sections, frozen sections, cell climbing slides, and cell smears for antibody stripping during TSA staining workflows \u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Experimental Procedure --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eEXPERIMENTAL PROCEDURE\u003c\/strong\u003e\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eSample Preparation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e1) Paraffin sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace slides sequentially into:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene I, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eXylene II, 15 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol I, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eAbsolute ethanol II, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e95% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003e85% ethanol, 5 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e75% ethanol, 5 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen rinse with distilled water.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e2) Frozen sections\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eFix frozen sections for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e3) Cell climbing slides or cell smears\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eFix cell samples for 10–30 min, wash with PBS for 5 min × 3, add 0.3% Triton X-100 permeabilization solution for 20 min, then wash with PBS for 5 min × 3.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"2\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eAntigen Retrieval\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePlace tissue sections into a retrieval container filled with either:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003epH 9.0 EDTA alkaline antigen retrieval solution, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003epH 6.0 citrate retrieval buffer\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003ePerform antigen retrieval in a microwave oven (other methods such as\u003c\/p\u003e\n\u003cul\u003e\n\u003cli style=\"line-height: 1.15;\"\u003epressure cooker 1–2 min\u003c\/li\u003e\n\u003cli style=\"line-height: 1.15;\"\u003eboiling water at 100°C for 15 min\u003c\/li\u003e\n\u003cli style=\"line-height: 1.15;\"\u003ewater bath at 95°C for 20 min may also be used).\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eMicrowave program:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emedium heat for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003estop heating and stay still for 8 min\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emedium-low heat for 7 min\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eDuring this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter natural cooling, place slides into PBS (pH 7.4), and wash on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eThe choice of retrieval buffer and conditions depends on tissue type, fixation method, and antigen type. This step may be omitted for frozen sections and cell samples.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"3\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Endogenous Peroxidase\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eIncubate the slides in 3% H₂O₂ solution at room temperature, protected from light, for 15 min. Then wash in PBS (pH 7.4) on a shaker 3 times for 5 min each.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"4\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003eBlock Non-specific Binding\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAfter gently removing excess liquid, draw a hydrophobic barrier around the tissue with a PAP pen.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd antibody diluent (or another blocking reagent such as 3% BSA or goat serum) to completely cover the tissue and block at room temperature for 30 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The antibody diluent contains various stabilizers and preservatives and can be used for blocking or for dilution of primary antibodies. Diluted primary antibodies can be stored long-term at 4°C and for up to one month at room temperature.\u003c\/p\u003e\n\u003col style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\" start=\"5\"\u003e\n\u003cli style=\"margin: 0 0 4px 0; background: transparent !important;\"\u003ePrimary Antibody Incubation\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eGently remove the blocking solution and add the appropriately diluted primary antibody working solution to the slide. Incubate flat in a humidified chamber protected from light:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eovernight at 4°C, or\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003e1–2 h at 37°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd a small amount of water into the humid chamber to prevent antibody evaporation.\u003c\/p\u003e\n\u003cp\u003e    6. Secondary Antibody Incubation\u003c\/p\u003e\n\u003cp\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add the HRP-conjugated secondary antibody corresponding to the host species of the primary antibody to fully cover the tissue. Incubate at room temperature for 50 min, protected from light, then wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The kit includes a ready-to-use HRP goat anti-rabbit\/mouse universal secondary antibody with very high sensitivity. No preparation is required.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e     7. Fluorescence Staining\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eAdd the diluted TSA fluorophore working solution to completely cover the tissue and incubate at room temperature for 1–15 min.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eRecommended reaction time: 5–10 min\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eThen wash with PBS 3 times for 5 min each.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e In a pilot experiment, stain for 1 min first, wash, and observe the result. If the positive signal is weak, continue applying the fluorophore reaction solution until suitable intensity is achieved before moving to the next step.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003e     8. Antibody Stripping\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp dir=\"ltr\"\u003e\u003cspan\u003eAdd an appropriate amount of \u003c\/span\u003e\u003cstrong\u003emIHC Antibody Stripping Buffer\u003c\/strong\u003e\u003cspan\u003e, prewarmed to \u003c\/span\u003e\u003cspan\u003e37°C until fully dissolved\u003c\/span\u003e\u003cspan\u003e, to cover the sample (\u003c\/span\u003e\u003cspan\u003erecommended for cell climbing slides, cell smears, frozen sections, and bone tissue prone to detachment\u003c\/span\u003e\u003cspan\u003e).\u003c\/span\u003e\u003c\/p\u003e\n\u003cul\u003e\n\u003cli dir=\"ltr\" aria-level=\"1\"\u003e\n\u003cp dir=\"ltr\" role=\"presentation\"\u003e\u003cstrong\u003eIncubate at 37°C for 5–20 min \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli dir=\"ltr\" aria-level=\"1\" style=\"font-weight: bold;\"\u003e\n\u003cp dir=\"ltr\" role=\"presentation\"\u003e\u003cstrong\u003eDiscard the stripping buffer \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli dir=\"ltr\" aria-level=\"1\" style=\"font-weight: bold;\"\u003e\n\u003cp dir=\"ltr\" role=\"presentation\"\u003e\u003cstrong\u003eAdd fresh stripping buffer again to cover the sample \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli dir=\"ltr\" aria-level=\"1\" style=\"font-weight: bold;\"\u003e\n\u003cp dir=\"ltr\" role=\"presentation\"\u003e\u003cstrong\u003eIncubate at 37°C for 5–20 min \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli dir=\"ltr\" aria-level=\"1\" style=\"font-weight: bold;\"\u003e\n\u003cp dir=\"ltr\" role=\"presentation\"\u003e\u003cstrong\u003eDiscard the stripping buffer \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli dir=\"ltr\" aria-level=\"1\" style=\"font-weight: bold;\"\u003e\n\u003cp dir=\"ltr\" role=\"presentation\"\u003e\u003cstrong\u003eWash with PBS for 5 min, repeat 3 times \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 6px 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003eParaffin sections may use either heat retrieval stripping or stripping buffer.\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003eCells and frozen sections must use the mIHC-specific antibody stripping buffer.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003e     9. Second-round Labeling\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eSwitch to another TSA fluorophore working solution and repeat Steps 3–7.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e     10. DAPI Nuclear Counterstaining\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, add ready-to-use DAPI staining solution within the marked area and incubate at room temperature for 5–20 min, protected from light.\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e      11. Mounting\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003eWash slides in PBS (pH 7.4) on a shaker 3 times for 5 min each. After gently removing excess liquid, mount with the anti-fade mounting medium.\u003c\/p\u003e\n\u003cp\u003e      12. Microscope and Imaging\u003c\/p\u003e\n\u003cp style=\"margin: 0 0 4px 0; line-height: 1.15; background: transparent !important;\"\u003eObserve and acquire images using:\u003c\/p\u003e\n\u003cul style=\"margin: 0 0 0 18px; padding-left: 14px; line-height: 1.15; background: transparent !important;\"\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003efluorescence microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003econfocal microscope\u003c\/li\u003e\n\u003cli style=\"margin: 0 0 3px 0; background: transparent !important;\"\u003emultichannel fluorescence scanner\u003c\/li\u003e\n\u003cli style=\"margin: 0; background: transparent !important;\"\u003emultispectral imaging system\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Precautions --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0 0 10px 0;\"\u003e\n\u003cp style=\"margin: 0 0 6px 0; line-height: 1.15; background: transparent !important;\"\u003e\u003cstrong\u003ePRECAUTIONS\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eThe stripping efficiency depends on section thickness, stripping temperature and time, and the type of primary antibody. The exact stripping time should be adjusted according to the specific situation. \u003c\/span\u003e\u003c\/li\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eIf antibody stripping is problematic, you may appropriately reduce the primary antibody concentration or place those antibodies in the final staining round 2.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eThe antibody stripping buffer can flow off the sample easily, so if runoff is observed it should be replenished promptly. \u003c\/span\u003e\u003c\/li\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eBefore each use, ensure the reagent is fully dissolved with no precipitate. If precipitation is present, place it at 37°C until fully dissolved before use. \u003c\/span\u003e\u003c\/li\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eThe mIHC-specific antibody stripping buffer is primarily recommended for cell climbing slides, cell smears, frozen sections, and bone tissue prone to section loss. For paraffin sections, EDTA or citrate retrieval buffer is preferred for antibody stripping. \u003c\/span\u003e\u003c\/li\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eThe mIHC-specific antibody stripping buffer is slightly acidic. Over-stripping (too long or too high a temperature) may lead to reduced antigen recognition and weaker DAPI nuclear staining. \u003c\/span\u003e\u003c\/li\u003e\n\u003cli dir=\"ltr\" role=\"presentation\"\u003e\u003cspan\u003eUnfixed sections are not suitable for use with this mIHC-specific antibody stripping buffer. \u003c\/span\u003e\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cb id=\"docs-internal-guid-d300df20-7fff-83d8-8cfc-803e33308f0e\"\u003e\u003c\/b\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003c!-- Notes --\u003e\u003c\/p\u003e\n\u003cdiv style=\"background: #efeff4; padding: 14px 16px; margin: 0;\"\u003e\n\u003cp style=\"margin: 0; line-height: 1.15; background: transparent !important;\"\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cem\u003eFor research use only. Not for use in diagnostic or therapeutic procedures.\u003c\/em\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003cp\u003e\u003cbr\u003e\u003c\/p\u003e","brand":"Bioss","offers":[{"title":"30ml","offer_id":52949119598891,"sku":"C2208","price":150.0,"currency_code":"USD","in_stock":true}]}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/collections\/IHC_IF_reagents.jpg?v=1740607987","url":"https:\/\/www.biossusa.com\/collections\/ihc-if-reagents\/isotype-control.oembed","provider":"Bioss","version":"1.0","type":"link"}