Cell Signaling Technology

Product Pathways - Metabolism

Phospho-IRS-1 (Tyr1222) Antibody #3066

Applications Reactivity Sensitivity MW (kDa) Source
W R (H) (M) Transfected Only 180 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-IRS-1 (Tyr1222) Antibody detects transfected levels of IRS-1 only when phosphorylated at Tyr1222. The antibody may cross-react with other activated receptor tyrosine kinases (RTKs) and docking proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1222 of human IRS-1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from CHO cells stably transfected with IRS-1, untreated or stimulated with insulin, using Phospho-IRS-1 (Tyr1222) Antibody (upper) or IRS-1 Antibody #2382 (lower).

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).

Phosphorylation of tyrosine 1222 of IRS-1 was identified in insulin stimulated cells (11). Phosphorylated Tyr1222 provides a docking site for the SH2 domain of PTP2C, which may mediate dephosphorylation of IRS-1 and lead to negative feedback of insulin signaling (12).

  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.
  11. Sun, X.J. et al. (1993) Mol. Cell. Biol. 13, 7418-7428.
  12. Rocchi, S. et al. (1995) Endocrinology 136, 5291-5297.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products