IRS-1 (Phospho-Ser323)Colorimetric Cell-Based ELISA Kit

This cell-based ELISA kit from Assay Genie is designed to measure phosphorylation of IRS-1 at serine 323 in cultured cells. It is described as a convenient, lysate-free, high-throughput and sensitive assay that uses an indirect ELISA format: IRS-1 (Phospho-Ser323) is captured by phospho-specific primary antibodies and detected with HRP-conjugated secondary antibodies, which catalyse a colorimetric reaction read at 450 nm.

Reactivity is reported for Human, Mouse and Rat, and the assay operates with a dynamic range of > 5000 Cells. The kit is suited to adherent cell cultures and can be used both to determine relative levels of phosphorylated IRS-1 and to screen the effects of treatments, siRNA or chemical inhibitors and activators on IRS-1 phosphorylation. Normalisation is supported by a monoclonal anti-GAPDH antibody as an internal positive control, together with Crystal Violet whole-cell staining to account for plating and cell-density differences after the colorimetric read.

IRS-1 is a major substrate of the insulin receptor kinase and a central adaptor in insulin signalling. According to the manufacturer, phosphorylation of IRS-1 at serine 323 has been linked to insulin resistance and to metabolic disorders such as diabetes and obesity, making this phospho-specific tool relevant to studies of insulin signalling, glucose homeostasis and related disease mechanisms.