IRS (insulin receptor substrate) is a key mediator of insulin and IGF signaling and is important for cellular functions such as growth, survival, and metabolism. IRS proteins contain no intrinsic enzymatic activity, but rather contribute to signaling through their function as adaptor proteins that serve to recruit effector proteins, such as PI3K, Grb-2, SHP-2, Fyn, c-Crk, CrkII and Nck to tyrosine receptor kinases (IR, IGF1R, VEGFR, EGFR, etc.) as well as other receptors. IRS proteins play an important regulatory function and are associated with various metabolic diseases and cancer.IRS proteins are phosphorylated on multiple tyrosine sites which serve as docking sites for SH2 containing proteins. Furthermore, IRS signaling is down regulated by serine or threonine phosphorylation, an important mechanism of insulin resistance during acute injury and infection, or chronic stress associated with aging or obesity. IRS can be serine/threonine phosphorylated by various kinases such as Akt, PKC, GSK3, mTOR, JNK, p38, ERK, etc. under conditions of hyperinsulinemia, obesity, inflammation, excess cytokines, lipid accumulation, etc. For example, hyperinsulinemia can lead to phosphorylation of IRS1 on serine 636 by mTOR. Understanding the regulation and signaling by IRS in cell growth, metabolism and survival will lead to new strategies to prevent or cure diabetes and other metabolic diseases.There are four members of the IRS family. Humans express IRS1, IRS2 and IRS4 while rodents also express Irs3. IRS1 and IRS2 are ubiquitously expressed and are the primary mediators of mitogenic and metabolic effects of insulin/IGF in most cell types. IRS1 and IRS4 are most often associated with tumor growth and proliferation. IRS2 is most often associated with tumor motility and invasion. IRS4 is found primarily in brain, kidney, thymus and liver. The MILLIPLEX® MAP 2-Plex Phospho/Total IRS1 kit detects total IRS1 and phosphorylated IRS1 (Ser636).The MILLIPLEX® 2-Plex Phospho/Total IRS1 kit contains all the components necessary to measure total IRS1 and phosphorylated IRS1 (Ser636). The MILLIPLEX® 2-Plex Phospho/Total IRS1 magnetic bead kit has been developed for the simultaneous detection of phosphorylated IRS1 (Ser636) and total IRS1 in a single well using the Luminex system. This detection assay is a rapid, convenient alternative to Western Blotting and immunoprecipitation procedures for the analysis of cell lysate samples.
Intracellular Bead-Based Multiplex Assays using the Luminex technology enables the simultaneous relative quantitation of multiple phosphorylation or total pathway proteins in tissue and cell lysate samples. An overnight (4°C) incubation is recommended for best results.This assay requires 25 μL diluted, filtered cell lysate per well.This kit must be run using the provided Assay Buffer.The suggested working range of protein concentration for the assay is 1 to 25 μg of total protein/well (25 μL/well at 40 to 1,000 μg/mL)Analytes available:IRS1 (Total)IRS1 (Ser636)
Luminex is a registered trademark of Luminex Corp
MILLIPLEX is a registered trademark of Merck KGaA, Darmstadt, Germany
xMAP is a registered trademark of Luminex Corp
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.