Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Antibody Pair #7288

Kit Includes Volume Cap Color
IRS-1 Mouse Capture Antibody (100X) 0.4 milliliters Pink
Phospho-IRS-1 (Ser307) Rabbit Detection Antibody (100X) 0.4 milliliters Blue
Anti-Rabbit IgG, HRP-Linked Antibody (1000X) 0.04 milliliters Red

Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human

Description

CST's PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Kit #7287. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The IRS-1 Mouse Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysate is added followed by a Phospho-IRS-1 (Ser307) Rabbit Detection Antibody and an HRP-conjugated, Anti-Rabbit IgG Antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of IRS-1 phosphorylated at Ser307.*Antibodies in this kit are custom formulations specific to the kit.

Sandwich ELISA

Sandwich ELISA

The relationship between the protein concentration of the lysate from untreated and insulin-treated hSkMC cells and the absorbance at 450 nm using PathScan® Phospho-IRS-1 (Ser307) Sandwich ELISA Antibody Pair #7288 is shown. After overnight starvation, hSkMC cells were treated with insulin (100 nM) for 15 minutes at 37°C and then lysed.

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 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, which suggests a potential mechanism for insulin resistance in some models of obesity (10).

  1. Sun, X.J. et al. (1991) Nature 352, 73-77.
  2. Sun, X.J. et al. (1992) J. Biol. Chem. 267, 22662-22672.
  3. Myers Jr., M.G. et al. (1993) Endocrinology 132, 1421-1430.
  4. Wang, L.M. et al. (1993) Science 261, 1591-1594.
  5. Rui, L. et al. (1997) J. Clin. Invest. 107, 181-189.
  6. Gao, Z. et al. (2002) J. Biol. Chem. 277, 48115-48121.
  7. Horike, N. et al. (2003) J. Biol. Chem. 278, 18440-18447.
  8. Ozes, O.N. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4640-4645.
  9. De Fea, K. and Ruth, R.A. (1997) Biochemistry 36, 12939-12947.
  10. Li, Y. et al. (2004) J. Biol. Chem. 279, 45304-45307.

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