IRS-1 (Phospho-Ser307)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed for the sensitive and accurate detection of phosphorylated IRS-1 at serine 307 in cell lysates. It targets IRS-1 (Phospho-Ser307) with reactivity to Human, Mouse, and Rat samples, and uses a colorimetric detection method measured at 450 nm. The kit is described as a convenient, lysate-free, high-throughput and sensitive assay that can detect IRS-1 protein phosphorylation and expression profile in cells, and can be applied to measuring the relative amounts of phosphorylated IRS-1 in cultured cells as well as screening the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on IRS-1 phosphorylation.

Detection is achieved through an indirect ELISA format, in which IRS-1 (Phospho-Ser307) is captured by phospho-specific primary antibodies and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalizing target absorbance values, and a Crystal Violet whole-cell staining method may be used to determine cell density; following staining, results can be analysed by normalizing absorbance values to cell amounts. The stated dynamic range is greater than 5000 cells.

Phosphorylation of IRS-1 at serine 307 has been implicated in insulin resistance, a hallmark of type 2 diabetes and other metabolic disorders, positioning this kit as a tool for studying insulin signaling pathways and regulation of glucose metabolism. Database information lists the target as IRS1, with Gene ID 3667 and UniProt ID P35568.