The Sphingosine 1 Phosphate （S1P） BioAssay ELISA Kit is a quantitative competitive assay for the detection of S1P in human serum，plasma，tissue homogenates and other biological fluids.Detection Range：3.125-200ng/mlSensitivity：<1.875ng/mlPrecision：Intra-assay CV： <8%Inter-assay CV： <10%Assay Principle：This ELISA kit employs the quantitative competitive enzyme-linked immunoassay technique，utilizing a microtiter plate provided in this kit has been pre-coated with S1P. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to S1P. 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 S1P，Biotin-conjugated antibody and enzyme-conjugated Streptavidin 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 S1P in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components：*370208A： Microtiter Strips，1x96 wells （8x12 wells）.*370208B： Standard，2x1 vials370208C： Sample/Standard Dilution Buffer，1x20ml370208D： Antibody （Biotin） （Concentrated），1x60ul370208E： Antibody Dilution Buffer，1x10ml370208F： Streptavidin （HRP） （SABC），1x120ul370208G： SABC Dilution Buffer，1x10ml370208H： TMB Substrate，1x10ml370208J： Stop Solution，1x10ml370208K： Wash Buffer，25X，1x30mlStorage and Stability：Store unopened *370208A at 4ºC; store at -20ºC once opened. Store unopened *370208B at 4°C; once reconstituted，store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store 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 and samples to wells.2. Add 50ul standard or sample to each well.3. Immediately add 50ul Antibody-Biotin working solution to each well and incubate for 45 minutes at 37°C4. Aspirate and wash 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate and wash 5 times.7. Add 90ul TMB substrate. Incubate 15-20 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.
