Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-Src Family (Tyr416) Antibody #2101

Applications Reactivity Sensitivity MW (kDa) Source
W H M R (C) (X) Endogenous 60 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  C=Chicken  X=Xenopus
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

Phospho-Src Family (Tyr416) Antibody detects endogenous levels of Src only when phosphorylated at tyrosine 416. The antibody may cross-react with other Src family members (Lyn, Fyn, Lck, Yes and Hck) when phosphorylated at equivalent sites. It does not cross-react with Src phosphorylated at tyrosine 527. It may cross react with phosphorylated RTKs.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide corresponding to residues surrounding Tyr416 of human Src. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-starved COLO 201 cells, v-Src antibody immunoprecipitates from COLO 201 cells and PDGF-treated NIH/3T3 cells, treated with lambda phosphatase as indicated, using Phospho-Src (Tyr416) Antibody (upper) or v-Src antibody (lower).

Background

The Src family of protein tyrosine kinases (including Src, Lyn, Fyn, Yes, Lck, Blk and Hck) are important in the regulation of growth and differentiation of eukaryotic cells (1). Src activity is regulated by tyrosine phosphorylation at two sites, but with opposing effects. Phosphorylation of Tyr416 in the activation loop of the kinase domain by Csk upregulates enzyme activity, whereas phosphorylation of Tyr527 in the carboxy-terminal tail renders the enzyme less active (2).

Lyn is a member of the Src family that is predominantly expressed in hematopoietic cells (3). Lyn participates in signaling from multiple cell surface receptors such as the B cell antigen receptor (BCR) and CD40 (4).Lck is essential for T-lymphocyte activation and differentiation (5,6). Phosphorylation of the carboxy-terminal Tyr505 downregulates Lck activity, while phosphorylation at Tyr394 leads to an increase in Lck activity (7).

  1. Thomas, S.M. and Brugge, J.S. (1997) Annu. Rev. Cell Dev. Biol. 13, 513-609.
  2. Hunter, T. (1987) Cell 49, 1-4.
  3. Yamanshi,, Y. et al. (1989) Proc. Natl. Acad. Sci. USA 86, 6538-6542.
  4. Yamanashi, Y. et al. (1991) Science 251, 192-194.
  5. Molina, T. J. et al. (1992) Nature 357, 161-164.
  6. Straus, D. B. et al. (1992) Cell 70, 585-593.
  7. Chow, L. M. et al. (1993) Nature 365, 156-160.

Application References

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Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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