Cystathionine Gamma Lyase (CSE) BioAssay(TM) ELISA Kit (Rat) is a sandwich enzyme immunoassay for in vitro quantitative measurement of CSE in rat tissue homogenates and other biological fluids. Detection Range: 0.156-10ng/ml Sensitivity: 0.053ng/ml est Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to CSE. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to rat CSE. 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 CSE, 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 CSE in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *024516A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *024516B: Standard, 2x1vial 024516C: Standard Diluent, 1x20ml *024516D: Detection Reagent A, 1x120ul *024516E: Detection Reagent B, 1x120ul 024516F: Assay Diluent A (2x),1x6ml 024516G: Assay Diluent B (2x), 1x6ml 024516H: TMB Substrate, 1x9ml 024516K: Stop Solution, 1x6ml 024516L: Wash Buffer, 30X, 1x20ml Storage and Stability: Store *024516A, *024516B, *024516D and *024516E 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 2 hours at 37C. 3. 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 15-25 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.
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