Product Pathways - Glucose Metabolism
Phospho-IRS-1 (Ser302) Antibody #2384
| Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|
| W | H R (M) (Pg) | 180 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
H=Human
M=Mouse
R=Rat
Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-IRS-1 (Ser 302) Antibody detects endogenous levels of IRS-1 only when phosphorylated at Ser302 of mouse IRS-1 or Ser307 of human IRS-1. This antibody does not detect IRS-1 phosphorylated at other sites.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser 302 of mouse IRS-1. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from CHO IR/IRS-1 cells ( transfected with insulin receptor and IRS-1), untreated or insulin-treated (100 nM for 5 min), showing an increase in phospho-IRS-1 (Ser302) with insulin stimulation, using Phospho-IRS1 (Ser302) Antibody. To confirm phospho-specificity of the antibody, a duplicate membrane was treated with calf intestinal alkaline phosphatase (CIP) after Western transfer.
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).
Ser302 in rat/mouse IRS-1 (corresponding to Ser307 of human IRS-1) is phosphorylated rapidly during insulin stimulation and has a postive role in IRS-1 tyrosine phosphorylation. Inhibition of Ser302 phosphorylation by short-term amino acid/glucose starvation correlates with a decrease in IRS-1 tyrosine phosphorylation without inhibition of insulin receptor autophosphorylation or Akt phosphorylation. A defect in this positive regulatory pathway may be a mechanism contributing to insulin resistence (11).
- Sun, X.J. et al. (1991) Nature 352, 73-77.
- Sun, X.J. et al. (1992) J. Biol. Chem. 267, 22662-22672.
- Myers Jr., M.G. et al. (1993) Endocrinology 132, 1421-1430.
- Wang, L.M. et al. (1993) Science 261, 1591-1594.
- Rui, L. et al. (1997) J. Clin. Invest. 107, 181-189.
- Gao, Z. et al. (2002) J. Biol. Chem. 277, 48115-48121.
- Horike, N. et al. (2003) J. Biol. Chem. 278, 18440-18447.
- Ozes, O.N. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4640-4645.
- De Fea, K. and Ruth, R.A. (1997) Biochemistry 36, 12939-12947.
- Li, Y. et al. (2004) J. Biol. Chem. 279, 45304-45307.
- White, M. F. et al. (2004) J. Biol. Chem. 279, 3447-3454.
Application References
- Tzatsos, A. and Kandror, K.V. (2006) Mol Cell Biol 26, 63-76. This article references the use of Phospho-IRS-1 (Ser302) Antibody in the following applications: Western Blotting
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Companion Products
- 2381 Phospho-IRS-1 (Ser307) Antibody
- 2382 IRS-1 Antibody
- 2386 Phospho-IRS-1 (Ser612) Antibody
- 2388 Phospho-IRS-1 (Ser636/639) Antibody
- 2389 Phospho-IRS-1 (Ser789) Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 2390 IRS-1 (59G8) Rabbit mAb
- 3193 Phospho-IRS-1 (Ser612) (L7B8) Mouse mAb
- 3194 IRS-1 (L3D12) Mouse mAb