KSR (Phospho-Ser392)Colorimetric Cell-Based ELISA Kit

Designed for the precise detection of phosphorylated KSR (Kinase Suppressor of Ras) protein at Ser392 in cell lysates, this Colorimetric Cell-Based ELISA Kit from Assay Genie is a lysate-free, high throughput and sensitive assay. It can be used to measure the relative amounts of phosphorylated KSR in cultured cells, as well as to screen for the effects that various treatments, inhibitors or activators have on KSR phosphorylation. Qualitative determination of KSR (Phospho-Ser392) concentration is achieved by an indirect ELISA format, in which the target is captured by phospho-specific primary antibodies while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyse a colorimetric reaction upon substrate addition.

Reactivity is established for Human and Mouse samples, with detection read at 450 nm and a dynamic range of > 5000 Cells suitable for adherent cells. Two normalisation approaches are incorporated: a monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalising the target absorbance values, and Crystal Violet whole-cell staining may be used to determine cell density following the colorimetric measurement, allowing absorbance values to be normalised to cell amounts and adjusting for plating differences.

KSR is described as a serine/threonine kinase of the Raf family functioning as a location-regulated scaffolding protein that connects MEK to RAF and promotes MEK and RAF phosphorylation and activity through assembly of an activated signalling complex. Phosphorylation of KSR at Ser392 plays a role in cell signalling pathways, particularly in the regulation of cell growth and proliferation, and dysregulation has been linked to cancer and inflammatory disorders, making it a valuable biomarker for studying these conditions. The target corresponds to gene symbol KSR1 (Gene ID: 8844, UniProt ID: Q8IVT5), with a reported molecular weight of 84,697 Da.