The Angiotensin 1-7 (Ang1-7) BioAssay ELISA Kit is a quantitative sandwich assay for the detection of Ang1-7 in rat serum,plasma,tissue homogenates and other biological fluids.Detection Range:7.813-500pg/mlSensitivity:<4.688pg/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 Ang1-7. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to Ang1-7. 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 Ang1-7,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 Ang1-7 in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components:*369731A: Microtiter Strips,1x96 wells (8x12 wells)*369731B: Standard,2x1 vial369731C: Sample/Standard Dilution Buffer,1x20ml369731D: Antibody-Biotin (Concentrated),1x120ul369731E: Antibody Dilution Buffer,1x10ml369731F: Streptavidin-HRP Conjugate (SABC),1x120ul369731G: SABC Dilution Buffer,1x10ml369731H: TMB Substrate,1x10ml369731J: Stop Solution,1x10ml369731K: Wash Buffer,25X,1x30mlStorage and Stability:Store unopened *369731A at 4ºC; store at -20ºC once opened. Store unopened *369731B 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,samples and control (zero) to wells.2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C. Aspirate and wash 2 times.3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 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 15-30 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.