SGK (Phospho-Ser422)Colorimetric Cell-Based ELISA Kit

A Colorimetric Cell-Based ELISA Kit from Assay Genie is designed to measure levels of SGK (Serum and Glucocorticoid-Regulated Kinase) phosphorylated at Ser422 in cell lysates from adherent cells. This cell-based, lysate-free assay demonstrates reactivity with Human, Mouse, and Rat samples and operates with a dynamic range of > 5000 Cells. Detection is achieved via a colorimetric reaction at 450 nm in a microplate format, employing HRP-conjugated secondary antibodies.

The kit uses an indirect ELISA approach: primary antibodies specific for SGK (Phospho-Ser422) capture the target, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, allowing the HRP enzyme to catalyze a colour-generating reaction upon substrate addition. Because the Cell-Based ELISA yields qualitative data, two normalization methods are provided. A monoclonal anti-GAPDH antibody serves as an internal positive control for normalizing target absorbance values, and Crystal Violet whole-cell staining can be used to determine cell density. The kit is suited to measuring relative amounts of phosphorylated SGK in cultured cells and to screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on SGK phosphorylation. Phosphorylation of SGK at Ser422 is known to play a role in diseases such as cancer and diabetes, making it a useful biomarker for studying disease mechanisms.