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PathScan® Phospho-IRS-1 (panTyr) Sandwich ELISA Kit #7133

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    Supporting Data

    REACTIVITY H M R
    Application Key:
    • ELISA-ELISA 
    Species Cross-Reactivity Key:
    • H-Human 
    • M-Mouse 
    • R-Rat 

    Product Information

    Product Description

    The PathScan® Phospho-IRS-1 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IRS-1 when tyrosine phosphorylated. An IRS-1 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, IRS-1 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-Tyrosine Mouse Detection Antibody is added to detect tyrosine phosphorylation of the captured IRS-1 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of IRS-1 phosphorylated on tyrosine.

    *Antibodies in kit are custom formulations specific to kit.

    Protocol

    Specificity / Sensitivity

    CST's PathScan® Phospho-IRS-1 (panTyr) Sandwich ELISA Kit #7133 detects IRS-1 when tyrosine phosphorylated. As shown in Figure 1, a significant induction of IRS-1 tyrosine phosphorylation can be detected in CHO (IR/IRS-1) cells following treatment with insulin using the Phospho-IRS-1 (panTyr) Sandwich ELISA Kit #7133. The level of total IRS-1 (phospho and nonphospho) remains unchanged as shown by Western analysis and by PathScan® Total IRS-1 Sandwich ELISA Kit #7328. This kit also detects tyrosine phosphorylated IRS-1 in insulin treated MCF7 cells and 3T3-L1 adipocytes.
    This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

    Species Reactivity:

    Human, Mouse, Rat

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