IR (Phospho-Tyr1361)Colorimetric Cell-Based ELISA Kit

A colorimetric cell-based ELISA kit from Assay Genie is designed for the detection of IR (Insulin Receptor) phosphorylation at tyrosine 1361 in cell-based assays. The kit enables measurement of relative amounts of phosphorylated IR in cultured cells and screening of the effects of various treatments, inhibitors (e.g., siRNA or chemicals), or activators on IR phosphorylation. It is reactive with human, mouse, and rat samples and is suitable for adherent cell samples. Insulin receptor phosphorylation at this site plays a crucial role in insulin signalling, regulating glucose metabolism, cell growth, and differentiation, with dysregulation implicated in insulin resistance, diabetes, and other metabolic disorders.

The assay uses an indirect ELISA format in which IR (Phospho-Tyr1361) is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction at 450 nm upon substrate addition. The stated dynamic range is greater than 5000 cells. 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 may subsequently be used to determine cell density and adjust for plating differences. The target gene is INSR (Gene ID: 3643, UniProt ID: P06213), a receptor tyrosine kinase that mediates the pleiotropic actions of insulin through the PI3K-AKT/PKB pathway (responsible for most metabolic actions) and the Ras-MAPK pathway, which regulates gene expression and cooperates with the PI3K pathway to control cell growth and differentiation.