For the quantitative determination of human cholesterol 7alpha-Hydroxylase (CYP7A) in serum, plasma, tissue homogenates, and cell lysates. Sensitivity: 4.7pg/ml Detection Range: 18.75-1200pg/ml Specificity: This assay has high sensitivity and excellent specificity for detection of human CYP7A. No significant cross-reactivity or interference between human CYP7A and analogues was observed. Assay Principle: This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for CYP7A has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CYP7A present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CYP7A is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CYP7A bound in the initial step. The color development is stopped and the intensity of the color is measured. Kit Components: Microtiter Strips, 1x96 wells Standard (Freeze dried), 2xvial Biotin-antibody (100X), 1x120ul HRP-avidin (100X), 1x120ul Biotin-antibody Diluent, 1x15ml HRP-avidin Diluent, 1x15ml Sample Diluent, 1x50ml Wash Buffer (25X), 1x20ml TMB Substrate, 1x10ml Stop Solution: 1x10ml Storage and Stability: Store all kit components at 4C. Stable for 6 months after receipt. Once opened, the kit is stable for 1 month providing this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
