Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-GRB10 (Ser476) (D4E6) Rabbit mAb #11817

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M (R) Endogenous 80 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
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-GRB10 (Ser476) (D4E6) Rabbit mAb recognizes endogenous levels of GRB10 protein only when phosphorylated at Ser476.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser476 of human GRB10 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from serum starved NIH/3T3 cells, untreated or treated with insulin (100 nM, 5 min), hIGF-1 #8917 (100 ng/ml, 5 min), or λ phosphatase, using Phospho-GRB10 (Ser476) (D4E6) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

IP

IP

Immunoprecipitation of phospho-GRB10 (Ser476) from extracts of serum starved NIH/3T3 cells treated with insulin (100 nM, 5 min), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-GRB10 (Ser476) (D4E6) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-GRB10 (Ser476) (D4E6) Rabbit mAb.

Background

The GRB7 family of adaptor proteins consist of GRB7, GRB10 and GRB14, which all contain an amino-terminal proline-rich SH3 binding domain, followed by PH, PBS, and SH2 domains. Each member of the family has several splice variants (1). It has been reported that GRB10 interacts with many receptor tyrosine kinases (RTKs) as well as downstream signal molecules including Raf, Akt, and Nedd4 (1,2). Although it was originally thought that GRB10 is exclusively phosphorylated at serine residues (3), Src kinase family members have been shown to phosphorylate GRB10 at Tyr67 (4).

Recently GRB10 was shown to be a direct substrate of mTORC1 (5). It is phosphorylated by mTORC1 at Ser150, Ser428, and Ser476 upon insulin stimulation (5).

  1. Han, D. C. et al. (2001) Oncogene 20, 6315-6321.
  2. Jahn, T. et al. (2002) Mol. Cell. Biol. 22, 979-991.
  3. Ooi, J. et al. (1995) Oncogene 10, 1621-1630.
  4. Langlais, P. et al. (2000) Oncogene 19, 2895-2903.
  5. Hsu, P.P. et al. (2011) Science 332, 1317-22.

Application References

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

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