Cytochrome P450 24A1 Colorimetric Cell-Based ELISA

A Cell-Based ELISA kit from Assay Genie designed for the qualitative detection of Cytochrome P450 24A1 (CYP24A1) protein expression in cultured cells. The assay uses an indirect ELISA format in which target-specific primary antibodies are captured and detected with HRP-conjugated secondary antibodies, producing a colorimetric readout at 450 nm. The kit is lysate-free and suited to high-throughput work in a 96-Well Microplate format, with a dynamic range of > 5000 Cells. It can be used to measure the relative amounts of CYP24A1 in cells and to screen the effects of treatments, inhibitors (such as siRNA or chemicals) or activators on CYP24A1 expression.

Normalization is supported by two methods: a monoclonal anti-human GAPDH antibody supplied as an internal positive control, and Crystal Violet whole-cell staining to determine cell density after the colorimetric measurement, allowing absorbance values to be normalized to cell amounts so that plating differences can be adjusted. The kit is reactive with human samples and is stored at 4°C for 6 months. CYP24A1 is a mitochondrial cytochrome P450 enzyme (Gene ID: 1591; UniProt ID: Q07973) that catalyses the NADPH-dependent 24-hydroxylation of calcidiol and calcitriol, playing a central role in vitamin D catabolism and calcium homeostasis. Its dysregulation has been linked to disorders including hypercalcemia infantile, as well as to cardiovascular diseases and autoimmune conditions.