Cytochrome P450 4B1 Colorimetric Cell-Based ELISA

From Assay Genie, this Cell-Based ELISA Kit is designed for the detection of Cytochrome P450 4B1 levels in cell lysates and tissue homogenates. The colorimetric assay uses an indirect ELISA format in which target-specific primary antibodies capture Cytochrome P450 4B1, and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition, read at 450 nm. Described as offering high sensitivity and specificity, the kit is a convenient, lysate-free, high-throughput method suited to measuring relative protein expression in cultured cells and to screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on Cytochrome P450 4B1.

Two normalization approaches are included: a monoclonal antibody specific for human GAPDH serves as an internal positive control for target absorbance values, and a Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be normalized to cell amounts. The assay is intended for adherent cells and is formatted for a 96-well microplate, with reactivity for human samples and a stated dynamic range of > 5000 Cells. Storage conditions are 4°C for 6 months. The target, CYP4B1 (UniProt P13584, Gene ID 1580), is a heme-thiolate monooxygenase of the cytochrome P450 family that participates in an NADPH-dependent electron transport pathway and oxidizes a variety of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics, supporting its use in studies of drug metabolism, drug development, and toxicology.