The Renin (REN) BioAssay(TM) ELISA Kit (Mouse) is a sandwich enzyme immunoassay for the in vitro quantitative measurement of REN in mouse serum, plasma and other biological fluids. Detection Range: 3.12 - 200pg/ml Sensitivity: <1.31pg/ml Precision: Intra-Assay: CV<10% Inter-Assay: CV<12% Test Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to REN. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to REN. 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 REN, biotin-conjugated antibody and enzyme-conjugated Avidin 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 REN in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *027851A: Microtiter Plate, 1x96 wells. Pre-coated, ready to use. *027851B: Standard, 2x1vial 027851C: Standard Diluent, 1x20ml *027851D: Detection Reagent A, 1x120ul *027851E: Detection Reagent B, 1x120ul 027851F: Assay Diluent A, 1x12ml 027851G: Assay Diluent B, 1x12ml 027851H: TMB Substrate, 1x9ml 027851K: Stop Solution, 1x6ml 027851L: Wash Buffer, 30X, 1x20ml Storage and Stability: Store *027851A, *027851B, *027851D and *027851E at -20C. Store all the other components at 4C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap. Assay Procedure Summary: 1. Prepare all reagents, samples and standards. 2. Add 100ul standard or sample to each well. Incubate for 1 hour at 37C. 3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul TMB Substrate. Incubate 10-20 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.
