EGFR (Phospho-Ser1026)Colorimetric Cell-Based ELISA Kit

Assay Genie supplies a Cell-Based ELISA kit for the detection of EGFR phosphorylated at serine 1026, a receptor tyrosine kinase that participates in the control of cell growth and differentiation. The assay is intended for use with adherent cells and uses an indirect ELISA format in which EGFR (Phospho-Ser1026)-specific primary antibodies capture the target, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyse a colourimetric reaction upon substrate addition.

Reactivity has been validated for human, mouse and rat samples, with a dynamic range of > 5000 Cells and detection measured at 450 nm. The protocol is described as convenient and lysate-free, supporting high-throughput screening of the effects that various treatments, inhibitors (such as siRNA or chemicals) or activators have on EGFR phosphorylation.

Because the readout is qualitative and cell-based, two normalisation strategies are provided: a monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and Crystal Violet whole-cell staining can be used to determine cell density and adjust the absorbance readings for plating differences. The target protein corresponds to Gene ID 1956 and UniProt ID P00533.