The Leptin （LEP） BioAssay ELISA Kit （Sheep） is a quantitative sandwich assay for the detection of Leptin in sheep serum，plasma，tissue homogenates and other biological fluids.Detection Range：0.156-10ng/mlSensitivity：<0.094ng/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 Leptin. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to Leptin. 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 Leptin，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 Leptin in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components：*370073A： Microtiter Strips，8x12 wells.*370073B： Standard，2 vials370073C： Sample/Standard Dilution Buffer，1x20ml370073D： Antibody-Biotin （Concentrated），1x120ul370073E： Antibody Dilution Buffer，1x10ml*370073F： Streptavidin-HRP Conjugate （SABC），1x120ul370073G： SABC Dilution Buffer，1x10ml*370073H： TMB Substrate，1x10ml370073J： Stop Solution，1x10ml370073K： Wash Buffer （25X），1x30mlStorage and Stability：Store unopened *370073A at 4ºC; store at -20ºC once opened. Store unopened *370073B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store *370073F and *370073H 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.
