EGFR (Phospho-Ser695)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie is designed for the accurate detection of phosphorylated EGFR (Epidermal Growth Factor Receptor) at Serine 695 in cell samples, and is intended for use with adherent cell samples. The kit is reactive with human, mouse, and rat samples and uses a colorimetric detection method at 450 nm in a Cell-Based ELISA format, with a dynamic range of > 5000 Cells. It is a lysate-free, high-throughput assay that can detect EGFR protein phosphorylation and expression profiles in cells, making it suitable for measuring relative amounts of phosphorylated EGFR in cultured cells and for screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on EGFR phosphorylation.

Detection is achieved through an indirect ELISA format, in which EGFR (Phospho-Ser695) is captured by phospho-specific primary antibodies, with HRP-conjugated secondary antibodies binding the Fc region of the primary antibody to catalyse a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalising target absorbance values, and a Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalised to cell amounts so that plating differences can be adjusted. The target is classified as a Phospho sub-type specific to the EGFR gene (Gene ID: 1956; UniProt ID: P00533).