Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-Gab1 (Tyr627) (C32H2) Rabbit mAb #3233

Applications Reactivity Sensitivity MW (kDa) Isotype
W H (M) (R) Endogenous 110 Rabbit IgG

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-Gab1 (Tyr627) (C32H2) Rabbit mAb detects endogenous levels of Gab1 only when phosphorylated at Tyr627. The antibody may cross-react with phosphorylated Gab2 or Gab3, or with activated receptor tyrosine kinases (RTKs).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr627 of human Gab1.

Western Blotting

Western Blotting

Western blot analysis of cell extracts from T47D cells, untreated or stimulated with heregulin, using Phospho-Gab1 (Tyr627) (C32H2) Rabbit mAb (upper) or Gab1 Antibody #3232 (lower).

Background

The Grb-associated binder (Gab) family is a family of adaptor proteins recruited by a wide variety of receptor tyrosine kinases (RTKs) such as EGFR, HGFR, insulin receptor, cytokine receptor and B cell antigen receptors. Upon stimulation of RTKs by their cognate ligand, Gab is recruited to the plasma membrane where it is phosphorylated and functions as a scaffold (1-4). Multiple tyrosine phosphorylation sites of Gab1 protein have been identified (5). Phosphorylation of Tyr472 regulates its binding to p85 PI3 kinase (6,7). Phosphorylation of Gab1 at Tyr307, Tyr373 and Tyr407 modulates its association to PLCγ (8). Phosphorylation of Tyr627 and Tyr659 is required for Gab1 binding to and activation of the protein tyrosine phosphatase SHP2 (6,9).

  1. Holgado-Madruga, M. et al. (1996) Nature 379, 560-564.
  2. Weidner, K.M. et al. (1996) Nature 384, 173-176.
  3. Takahashi-Tezuka, M. et al. (1998) Mol. Cell. Biol. 18, 4109-4117.
  4. Ingham, R.J. et al. (2001) J Biol Chem 276, 12257-65.
  5. Lehr, S. et al. (1999) Biochemistry 38, 151-159.
  6. Rocchi, S. et al. (1998) Mol. Endocrinol. 12, 914-923.
  7. Yu, C.F. et al. (2001) J Biol Chem 276, 32552-8.
  8. Gual, P. et al. (2000) Oncogene 19, 1509-18.
  9. Cunnick, J.M. et al. (2001) J Biol Chem 276, 24380-7.

Application References

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