MDM2 (Phospho-Ser166)Colorimetric Cell-Based ELISA Kit

This cell-based ELISA kit from Assay Genie enables qualitative detection of MDM2 protein phosphorylation at Ser166 in adherent cell samples. The assay uses an indirect ELISA format in which MDM2 (Phospho-Ser166) is captured by phospho-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyze a colorimetric reaction upon substrate addition. The kit is lysate-free, high throughput, and sensitive, and can be used to measure relative levels of phosphorylated MDM2 in cultured cells and to screen the effects of treatments, inhibitors such as siRNA or chemicals, or activators on MDM2 phosphorylation.

Reactive with human and mouse samples, the assay is read colorimetrically at 450 nm and has a dynamic range of > 5000 cells. A monoclonal anti-human GAPDH antibody is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining method is provided to determine cell density so that absorbance values can be normalised to cell amounts and plating differences adjusted. MDM2 is a nuclear-localised E3 ubiquitin-protein ligase that mediates ubiquitination of p53/TP53, leading to its degradation by the proteasome; phosphorylation of MDM2 at Ser166 is reported to affect its function and may play a role in cancer development and progression.