Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Gab Antibody Sampler Kit #9791

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-Gab1 (Tyr307) Antibody #3234 40 µl W H M 115 Rabbit
Phospho-Gab1 (Tyr627) (C32H2) Rabbit mAb #3233 40 µl W H (M) (R) 110 Rabbit IgG
Gab1 Antibody #3232 40 µl W IP H M R Mk 110 Rabbit
Phospho-Gab2 (Ser159) Antibody #3884 40 µl W H M R 98 Rabbit
Phospho-Gab2 (Tyr452) (C33G1) Rabbit mAb #3882 40 µl W H M 98 Rabbit IgG
Gab2 (26B6) Rabbit mAb #3239 40 µl W H M 90 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Gab1 Antibody detects endogenous levels of total Gab1 protein. Gab2 Antibody detects endogenous levels of total Gab2 protein. Each activation-state antibody detects endogenous levels of Gab1 or Gab2 phosphorylated at the specified sites. There may be some cross-reactivity with other phosphorylated proteins. Please consult individual product web pages for more details.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293, NIH/3T3-L1, C6 and COS cells, using Gab1 Antibody #3232.

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 #3233 (upper) or Gab1 Antibody #3232 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293 cells, untreated or treated with In1B for indicated times, using Phospho-Gab1 (Tyr307) Antibody #3234 (upper) or Gab1 Antibody #3232 (lower).


Western Blotting

Western Blotting

Western blot analysis of cell extracts from various cell lines, using Gab2 (26B6) Rabbit mAb #3239.

Western Blotting

Western Blotting

Western blot analysis of extracts from Baf3 cells stably transfected with a construct expressing FLT3, untreated or treated with FLT ligand (FL), using Phospho-Gab2 (Tyr452) (C33G1) Rabbit mAb #3882 (upper) or Gab2 (26B6) Rabbit mAb #3239 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from H-4-II-E cells, untreated, HGF-treated, insulin-treated or treated with lambda phosphatase after lysis, using Phospho-GAB2 (Ser159) Antibody #3884.


Description

The Gab Antibody Sampler Kit provides an economical means of evaluating levels of Gab1 and Gab2 total protein, as well phosphorylated Gab1 and Gab2 at the specified sites. The kit contains enough reagents to perform four western blot experiments per primary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr472 of Gab1, or phosphopeptides corresponding to residues surrounding Tyr307 of human Gab1 or Ser159 of human Gab2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.Monoclonal antibodies are produced by immunizing animals with recombinant human Gab2, or synthetic phosphopeptides corresponding to residues surrounding Tyr627 of human Gab1 or Tyr452 of human Gab2.

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).

Gab2, a recently identified docking protein of Gab family, contains a pleckstrin homology domain and potential binding sites for SH2 and SH3 domain-containing proteins. Gab2 has been shown to support growth, differentiation and function in a number of hematopoietic cells (10).

  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.
  10. Yi, Q. et al. (2005) Biochem Biophys Res Commun 337, 446-51.

Application References

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