Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
Principle of the Assay
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human DEFa4. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human DEFa4. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human DEFa4, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human DEFa4 in the samples is then determined by comparing the OD of the samples to the standard curve.
For use with: Serum, plasma, tissue homogenates, cell lysates, and other biological fluids.
For Research Use Only. Not for use in diagnostic procedures.
Target Information
| SWISS PROT | P12838 |
| SYNONYMS | DEF4; HP-4; HNP-4; Neutrophil defensin 4 |
Materials Supplied
| Kit Components | 96 Wells Quantity/Size |
|---|---|
| Pre-Coated Microplate | 48T: 6 strips x 8 wells; 96T: 12 strips x 8 wells |
| Standard (Lyophilized) | 48T: 1 vial; 96T: 2 vials |
| Biotinylated Antibody (100×) | 48T: 60 μL; 96T: 120 μL |
| Streptavidin-HRP (100×) | 48T: 60 μL; 96T: 120 μL |
| Standard/Sample Diluent Buffer | 48T: 10 mL; 96T: 20 mL |
| Biotinylated Antibody Diluent | 48T: 6 mL; 96T: 12 mL |
| HRP Diluent | 48T: 6 mL; 96T: 12 mL |
| Wash Buffer (25×) | 48T: 10 mL; 96T: 20 mL |
| TMB Substrate Solution | 48T: 6 mL; 96T: 10 mL |
| Stop Reagent | 48T: 3 mL; 96T: 6 mL |
| Plate Covers | 48T: 1 piece; 96T: 2 pieces |
Storage
| Storage | *If the kit is opened, Store the whole kit at 4°C. If the kit is not used up in 1 week. Store the Pre-Coated Microplate, Standard ,Biotinylated Antibody and Streptavidin-HRP at -20°C, the rest reagents at 4°C, please used up within 6 months. *If the kit is not opened, store the whole kit: 4°C(short time storage, valid for 6 months); -20°C (long-term storage, valid for 1 year). Avoid repeated freeze-thaw cycles. |
Performance Characteristics
| REPEATABILITY | Intra-assay Precision (Precision within an assay)：CV% < 8% Three samples of known concentration were tested twenty times on 1 plate to assess intra-assay precision. Inter-assay Precision (precision between assays)：CV% < 10% Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. |
| SENSITIVITY | The minimum detectable dose was 14pg/mL. |
| ASSAY RANGE | 31.25–2,000pg/mL |
| SPECIFICITY | This assay has high sensitivity and excellent specificity for detection of Human DEFa4. No significant cross-reactivity or interference between Human DEFa4 and analogues was observed. |