The Cytochrome P450 7A1 (CYP7A1) BioAssay™ ELISA Kit (Rat) is a sandwich enzyme immunoassay for the in vitro quantitative measurement of CYP7A1 in rat tissue homogenates, cell lysates and other biological fluids.
Detection Range:1.56-100ng/ml
Sensitivity:<0.54ng/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 CYP7A1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to CYP7A1. 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 CYP7A1, 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 CYP7A1 in the sample is then determined by comparing the O.D. of the sample to the standard curve.
Kit Components:*365925A: Microtiter Plate, 1x96 wells. Pre-coated; ready to use
Storage and Stability:Store *365925A, *365925B, *365925D and *365925E at -20°C. Store all the other components at 4°C. 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 1 hour at 37°C
3. Aspirate and add 100ul Detection Reagent A. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul Detection Reagent B. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 90ul TMB Substrate. Incubate 10-20 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
