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

This Fluorometric Cell-Based ELISA Kit from Assay Genie is designed to monitor phosphorylation of IRS-1 at Serine 307 in cultured cells. The lysate-free, high-throughput format allows qualitative measurement of phosphorylated IRS-1 (Phospho-Ser307) and is suited to screening the effects of treatments, inhibitors such as siRNA or chemicals, and activators on IRS-1 phosphorylation. Detection is by indirect ELISA, with the target captured by specific primary antibodies and reported using Dye 1- and Dye 2-conjugated secondary antibodies, with readings taken at Ex/Em 651/667 nm and 495/521 nm.

The kit reacts with Human, Mouse and Rat samples and is configured for the cell-based workflow, with a stated dynamic range of greater than 5000 cells. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising RFU values, and an antibody against the non-phosphorylated counterpart of IRS-1 (Phospho-Ser307) is also provided so that the RFU value for phosphorylated IRS-1 can be normalised to that for total IRS-1. Storage is at 4 °C with a stability of 6 months. Associated database references are Gene ID 3667 and UniProt ID P35568.

IRS-1 is an adaptor protein and one of the major substrates of the insulin receptor kinase. It contains multiple tyrosine phosphorylation motifs that serve as docking sites for SH2-domain-containing proteins including the p85 subunit of phosphatidylinositol 3-kinase and GRB-2, and is itself phosphorylated on serine/threonine residues by kinases including JNK and IKK.