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. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
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. While phosphorylation at Tyr416 in the activation loop of the kinase domain upregulates enzyme activity, phosphorylation at Tyr527 in the carboxy-terminal tail by Csk 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
- Nikolic, D.S. et al. (2011) Blood 118, 4841-52. Applications: Western Blotting
- Harr, M.W. et al. (2010) Cell Death Differ 17, 1381-91. Applications: Western Blotting
- Zaytseva, Y.Y. et al. (2012) Cancer Res 72, 1504-17. Applications: Western Blotting
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Companion Products
- 2123 Src (32G6) Rabbit mAb
- 2102 Non-phospho-Src (Tyr416) (7G9) Mouse mAb
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- 2732 Lyn Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 2108 Src Antibody
- 2109 Src (36D10) Rabbit mAb
- 2110 Src (L4A1) Mouse mAb
- 2731 Phospho-Lyn (Tyr507) Antibody
- 2751 Phospho-Lck (Tyr505) Antibody
- 2752 Lck Antibody
- 4023 Fyn Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.
