Product Information

Product Description

CST's PathScan^® Total IRS-1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected IRS-1 protein. An IRS-1 Mouse mAb #3194* has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-IRS-1 proteins are captured by the coated antibody. Following extensive washing, IRS-1 Rabbit mAb #2397* is added to detect both the captured phospho- and nonphospho-IRS-1 protein. Anti-rabbit IgG, HRP-linked Antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total IRS-1 protein.
* Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

CST's PathScan^® Total IRS-1 Sandwich ELISA Kit #7214 detects IRS-1 protein. As shown in Figure 1, using PathScan^® Phospho-IRS-1 (Ser612) ELISA Kit #7213, a significant induction of phospho-IRS-1 (Ser612) is detected in CHO-IR/IRS-1 cells treated with insulin. However, the levels of total IRS-1 (phospho and nonphospho) detected by PathScan^® Total IRS-1 Sandwich ELISA Kit #7214 remain unchanged. In Figure 3, Western blot analysis of protein captured in the IRS-1 mouse mAb #3194 coated microwell shows a single band corresponding to the IRS-1 protein. In 3T3 cells, IRS-1 protein also can be detected by this kit (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Insulin receptor substrate 1 (IRS-1) is one of the major substrates of the insulin receptor kinase (1). IRS-1 contains multiple tyrosine phosphorylation motifs that serve as docking sites for SH2-domain containing proteins that mediate the metabolic and growth-promoting functions of insulin (2-4). IRS-1 also contains over 30 potential serine/threonine phosphorylation sites. Ser307 of IRS-1 is phosphorylated by JNK (5) and IKK (6) while Ser789 is phosphorylated by SIK-2, a member of the AMPK family (7). The PKC and mTOR pathways mediate phosphorylation of IRS-1 at Ser612 and Ser636/639, respectively (8,9). Phosphorylation of IRS-1 at Ser1101 is mediated by PKCθ and results in an inhibition of insulin signaling in the cell, suggesting a potential mechanism for insulin resistance in some models of obesity (10).

Database Links

UniProt ID: P35568

Entrez-Gene Id: 3667

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.

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PathScan® Total IRS-1 Sandwich ELISA Kit #7214

Supporting Data