{"title":"Cell Assay","description":null,"products":[{"product_id":"ba00208","title":"Enhanced Cell Counting Kit- 8 (CCK-8)","description":"\u003ctable style=\"width: 750px;\" data-mce-fragment=\"1\"\u003e\n\u003ctbody data-mce-fragment=\"1\"\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 566px;\" data-mce-fragment=\"1\" colspan=\"2\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eCat. No. BA00208\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eGeneral Information\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003eEnhanced Cell Counting Kit- 8 (CCK-8) allows very convenient assays by utilizing the highly water-soluble tetrazolium salt WST-8.\u003cbr\u003e\u003cbr\u003eWST-8 produces a water-soluble formazan dye upon reduction in the presence of an electron carrier and is reduced by dehydrogenases in cells to give a yellow-colored product namely formazan(as shown below). \u003cbr\u003e\u003cbr\u003eThe kit allows sensitive colorimetric assays for the determination of the number of viable cells in cell proliferation and cytotoxicity assays. The amount of the formazan dye generated by the activity of dehydrogenases in cells is directly proportional to the number of living cells.\u003c\/p\u003e\n\u003cimg data-mce-fragment=\"1\" alt=\"\" src=\"https:\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/BA00208_dehydrogenases_480x480.png?v=1631307479\" data-mce-src=\"https:\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/BA00208_dehydrogenases_480x480.png?v=1631307479\"\u003e\u003cbr data-mce-fragment=\"1\"\u003e\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eAdvantages\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cul data-mce-fragment=\"1\"\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eLow toxicity to cells\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eOne-bottle, no premixing of components\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eNo organic solvents or isotopes required\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eNo harvesting, no washing, and no solubilization steps\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eMore sensitive than MTT, XTT, MTS, or WST-1\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eStorage\u003c\/strong\u003e\u003c\/p\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003eStore the kit at 4°C with protection from light for one year.\u003cbr\u003eStore the kit at -20°C with protection from light for two years.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eKit Contents\u003c\/strong\u003e\u003c\/p\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e1ml(100T) \/ 5ml(500T) \/ 6*5ml(3000T)\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003e\u003cspan style=\"text-decoration: underline;\"\u003eGeneral Protocols:\u003c\/span\u003e\u003cbr\u003e\u003cbr\u003eCell Number Determination\u003c\/strong\u003e\u003c\/p\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003col data-mce-fragment=\"1\"\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eCollect the cells needed for the experiment and then prepare wells that contain known numbers of viable cells.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eIt is recommended to preincubate adherent cells. After that add 10 μl of the CCK-8 solution to each well of the plate.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eIncubate the plate for 1-4 hrs in the incubator.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eMeasure the absorbance at 450 nm using a microplate reader. Prepare a calibration curve using the data obtained from the wells that contain known numbers of viable cells.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eUnder the completely consistent condition, the cell number of the unknown sample can be calculated according to the standard curve.\u003cbr data-mce-fragment=\"1\"\u003e\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eCell Viability Assay\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003col data-mce-fragment=\"1\"\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eInoculate cell suspension (above 5000 cells\/100 μl\/well) in a 96-well plate. The cells for the assay should enter into the logarithmic growth phase. The average incubation time to enter into this phase is from 24 hrs to 48 hrs. Please check cell databases to estimate the preincubation time.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eAdd 10μl of the CCK-8 solution to each well of the plate. Be careful not to introduce bubbles to the wells.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eIncubate the plate for 1-4 hrs in the incubator. However, the absorbance will differ between cell types even if the number of cells\/well and coloration time are the same. Set an appropriate incubation time to give a proportional relationship between the cell number and the absorbance.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eMeasure the absorbance at 450 nm using a microplate reader. To measure the absorbance later, add 10μl of 1% w\/v SDS or 0.1 M HCl to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 24 hrs.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eCell Proliferation and Cytotoxicity Assay\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003col data-mce-fragment=\"1\"\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eDispense 100μl of cell suspension in a 96-well plate at a density of 5*103-105 cells\/well. Pre-incubate the plate overnight or 24hrs in a humidified incubator.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eAdd various concentrations of the toxicant into the culture media in the plate.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eIncubate the plate for an appropriate length of time (e.g., 3, 6, 12, 24, or 48 hrs) in the incubator.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eAdd 10 μl of CCK-8 solution to each well of the plate. Be careful not to introduce bubbles to the wells.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eIncubate the plate for 1-4 hrs in the incubator. The incubation time varies by the type and number of cells in a well.\u003c\/li\u003e\n\u003cli data-mce-fragment=\"1\" data-mce-style=\"font-weight: 400;\"\u003eMeasure the absorbance at 450 nm using a microplate reader. To measure the absorbance later, add 10μL of 1% w\/v SDS 0.1 M HCl to each well, cover the plate and store it with protection from light at room temperature. Be sure to take a reading within 24 hrs after stopping the reaction.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eThe Formula to Calculate:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003eThe ratio of cell viability = [(ODt-ODb) \/ (ODc-ODb)] × 100%\u003cbr\u003e\u003cbr\u003eThe ratio of inhibition = [(ODc-ODt) \/ (ODc-ODb)] × 100%\u003cbr\u003e\u003cbr\u003eODt: The absorbance of the test group (with cell、culture Medium、CCK-8 and toxicant)\u003cbr\u003e\u003cbr\u003eODc: The absorbance of the control (with cell、culture Medium and CCK-8, without toxicant)\u003cbr\u003e\u003cbr\u003eODb: The absorbance of blank (with culture medium and CCK-8, without cell and toxicant)\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003eQuality verification:\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cimg src=\"https:\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/files\/BA00208_Quality_Verification-s_480x480.png?v=1631307409\" alt=\"\"\u003e\u003c\/p\u003e\n\u003cp data-mce-fragment=\"1\"\u003eCell line: SP2\/0、0-105\/100ul\u003cbr\u003e\u003cbr\u003eCulture Medium:DMEM,10%FBS;37°C,5% CO2\u003cbr\u003e\u003cbr\u003eIncubation: 1h\\ 2h\\ 3h\u003c\/p\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr data-mce-fragment=\"1\"\u003e\n\u003ctd style=\"width: 156.078px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e\u003cstrong\u003ePrecautions:\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 409.922px;\" data-mce-fragment=\"1\"\u003e\n\u003cp data-mce-fragment=\"1\"\u003e1. The number of cells in a well varies by the size of the cell, proliferation rate, and specific requirements of the experiment. The total number of cells is recommended to be 0-105 per well.\u003cbr\u003e\u003cbr\u003e2. Prepare the well for background measurement which contains all materials except for cells. Measure the background absorbance of the well at 450 nm, and then subtract the background absorbance from the absorbance of the sample well containing all materials and cells.\u003cbr\u003e\u003cbr\u003e3. Please add Cell Counting Kit-8 solution equal to 1\/10 the volume of the media (if the media is 100μl, add 10μl of solution).\u003cbr\u003e\u003cbr\u003e4. If the cell culture time is long and the medium color changes, remove the culture medium from cells and add new culture medium prior to adding CCK-8.\u003cbr\u003e\u003cbr\u003e5. In order to fully mix the cck-8 and reduce the residue of cck-8 on the pipette, it is recommended to dilute the cck-8 reagent with culture medium before sample addition.\u003cbr\u003e\u003cbr\u003e6. WST-8 may react with reducing agents to generate WST-8 formazan and give a false reading. Please check the background O.D. if reducing agents are used in cytotoxicity assays or cell proliferation assays. The absorbance from such dyes can be subtracted as a blank.\u003cbr\u003e\u003cbr\u003e7. This product is for R\u0026amp;D use only, not for drug, household, or other uses.\u003cbr\u003e\u003cbr\u003e8. For your safety and health, please wear lab clothes and disposable gloves.\u003c\/p\u003e\n\u003cbr data-mce-fragment=\"1\"\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e","brand":"Bioss","offers":[{"title":"5ml (500T)","offer_id":39526364479539,"sku":"BA00208-500T","price":49.0,"currency_code":"USD","in_stock":true},{"title":"6*5ml (3000T)","offer_id":39526364512307,"sku":"BA00208-3000T","price":200.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1223\/3460\/products\/BA00208dehydrogenases.png?v=1631306663"}],"url":"https:\/\/www.biossusa.com\/collections\/cell-assay\/cross-reactive-species_dog.oembed","provider":"Bioss","version":"1.0","type":"link"}