Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-Src Family (Tyr416) Antibody #2101

supporting products
compare antibodies
control extracts

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

2101:: Western Blotting

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 animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr416 of human Src. Antibodies are purified by protein A and peptide affinity chromatography.

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, which includes 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 Src at Tyr416 in the activation loop of the kinase domain by carboxy-terminal Src kinase (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).

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

Application References

Gonfloni, S. et al. (1999) Leucine 255 of Src couples intramolecular interactions to inhibition of catalysis. Nat. Struct. Biol. 6, 760-764. Applications: Western Blot
Gonfloni, S. et al. (2000) Crosstalk between the catalytic and regulatory domains allows bidirectional regulation of Src. Nat. Struct. Biol. 7, 281-286. Applications: Western Blot
Ibitayo, A. I. et al. (1998) Src kinase and PI 3-kinase as a transduction pathway in ceramide-induced contraction of colonic smooth muscle. Am. J. Physiol. 275, 705-711. Applications: Western Blot
Wu, W. et al. (2002) Src-dependent phosphorylation of the epidermal growth factor receptor on tyrosine 845 is required for zinc-induced Ras activation. J. Biol. Chem. 277 (27), 24252-24257. Applications: Western Blotting
Martinon, F. et al. (2002) The Inflammasome: A Molecular Platform Triggering Activation of Inflammatory Caspases and Processing of proIL-beta. Mol. Cell 10, 417-426. Applications: Western Blotting
Nagata, Y. et al. (2004) PTEN activation contributes to tumor inhibition by trastuzumab, and loss of PTEN predicts trastuzumab resistance in patients. Cancer Cell 6, 117-127. Applications: Western Blotting
White, D. E. et al. (2004) Targeted disruption of beta1-integrin in a transgenic mouse model of human breast cancer reveals an essential role in mammary tumor induction. Cancer Cell 6, 159-170. Applications: IC-IF Western Blotting
Wang, D. et al. (2000) Distinct effects of N-acetylcysteine and nitricoxide on angiotensin II-induced epidermal growth factor receptor phosphorylation and intracellular Ca(2+) levels. J. Biol. Chem. 275, 12223-12230. Applications: Western Blot

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

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.