Cytochrome P450 2C19 Colorimetric Cell-Based ELISA

Assay Genie's Cell-Based ELISA Kit uses a Colorimetric 450 nm detection method for the qualitative detection of Cytochrome P450 2C19 (CYP2C19) protein expression in cultured cells, cell lysates, and tissue homogenates. The assay is reactive with human samples, operates over a dynamic range of > 5000 Cells, and is supplied in a 96-Well Microplate format with storage at 4°C for 6 Months. It is designated as a Cell-Based ELISA in the Metabolism research area.

The indirect ELISA format captures Cytochrome P450 2C19 using target-specific primary antibodies, with HRP-conjugated secondary antibodies binding the Fc region of the primary antibody. A monoclonal antibody specific for human GAPDH serves as an internal positive control for normalizing target absorbance values. Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalized to cell amounts. The kit can be used to measure relative amounts of Cytochrome P450 2C19 in cultured cells and to screen for the effects of treatments, inhibitors such as siRNA or chemicals, or activators on the target.

Cytochrome P450 2C19 belongs to the cytochrome P450 superfamily of enzymes, which are monooxygenases catalyzing many reactions involved in drug metabolism and the synthesis of cholesterol, steroids, and other lipids. The protein localizes to the endoplasmic reticulum and metabolizes many xenobiotics, including mephenytoin, omeprazole, diazepam, and some barbiturates. Variations in CYP2C19 activity impact the effectiveness and safety of drugs such as clopidogrel and omeprazole, making it a biomarker in personalized medicine and pharmacogenomic research. The gene is located on chromosome 10q24, with Gene ID 1557 and UniProt ID P33261; the protein has a molecular weight of 55,931 Da.