The Interleukin 17 (IL-17) BioAssay ELISA Kit (Canine) is a quantitative sandwich assay for the detection of IL-17 in canine serum,plasma,tissue homogenates and other biological fluids.Detection Range:31.25 - 2,000pg/mlSensitivity:<18.75pg/mlPrecision: .Intra-Assay CV: <8%Inter-Assay CV: <10%Assay Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to IL-17. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to IL-17. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added,only those wells that contain IL-17,biotin-conjugated antibody and enzyme-conjugated streptavidin complex will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of IL-17 in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components:*370021A: Microtiter Strips,8x12 wells.*370021B: Standard,2 vials370021C: Sample/Standard Dilution Buffer,1x20ml370021D: Antibody-Biotin (Concentrated),1x120ul370021E: Antibody Dilution Buffer,1x10ml*370021F: Streptavidin-HRP Conjugate (SABC),1x120ul370021G: SABC Dilution Buffer,1x10ml*370021H: TMB Substrate,1x10ml370021J: Stop Solution,1x10ml370021K: Wash Buffer (25X),1x30mlStorage and Stability:Store unopened *370021A at 4ºC; store at -20ºC once opened. Store unopened *370021B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store *370021F and *370021H in the dark at 4°C. Store all other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product,centrifuge the original vials after thawing and prior to removing the cap.Assay Summary:1. Wash plate 2 times before adding standards,samples and control (zero) to wells.2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C 3. Aspirate the liquid and wash plate 3 times.3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C4. Aspirate the liquid and wash plate 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate the liquid and wash plate 5 times.7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.