CST's PathScan® Phospho-IRS-1 (Ser612) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected phospho-IRS-1 (Ser612) protein. A Phospho-IRS-1 (Ser612) Mouse mAb #3193* has been coated onto the microwells. After incubation with cell lysates, phospho-IRS-1 (Ser612) proteins are captured by the coated antibody. Following extensive washing, IRS-1 Rabbit mAb #2397* is added to detect the captured phospho-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 phospho-IRS-1 (Ser612) protein.
* Antibodies in kit are custom formulations specific to kit.
CST's PathScan® Phospho-IRS-1 (Ser612) Sandwich ELISA Kit #7213 detects phospho-IRS-1 (Ser612) protein. As shown in Figure 1, using this ELISA Kit #7213, a significant induction of phospho-IRS-1 (Ser612) is detected in CHO-IR/IRS-1 cells treated with insulin. The levels of total IRS-1 (phospho and nonphospho) detected by PathScan® Total IRS-1 Sandwich ELISA Kit #7214 remain unchanged. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
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