Src is a protein tyrosine kinase known to regulate cellular adhesion. Composed of three major domains; SH2, SH3 and a kinase catalytic domain, Src can be switched from an inactive to an active state though control of its phosphorylation state or through protein-protein interaction. Phosphorylation of Y529 by Csk and Chk inactivates Src, while dephosphorylation of Y529 will modify its conformation to an open active state (1-3) Mutation of Y529 leads to Scr overactivity. Several cancers including colon and breast cancer have been associated with an increase of Src activity (4).
Recommended Applications
WB, ICC, IP
Applications and Recommended Dilution Factors
WB
IHC
ICC
FC
IP
1:5,000-10,000
1:50
1:50
Species Reactivity*
Human
Mouse
Rat
Positive
Positive
Positive
*Cross reactivity determined by western blot only.
Product Data
A. Western blot analysis on A431 cell lysate using anti-Phospho-Src (pY529) RabMAb (cat. # 1550-1), dilution 1:10,000. Cells were either (A) untreated, (B) treated with EGF and (C) treated with EGF+ alkaline phosphatase.
B. Dot Blot analysis on immunogen phospho-peptide (A) and non-phospho peptide (B) using anti-Phospho-Src (pY529) RabMAb (cat. # 1550-1) dilution 1:2,000
C. Immunofluorescent staining of A431 cells using anti-Phospho-Src (pY529) RabMAb (cat. # 1550-1).
Specificity
A synthetic phospho-peptide corresponding to residues surrounding Tyrosine 529 of human Src was used as immunogen. The antibody will detect Src phosphorylation on Tyrosine 529 of both isoforms.
Storage Buffer & Conditions
50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA.
1. Cooper, J. A., and King, C. S. (1986) Mol. Cell. Biol. 6, 4467-4477 2. Courtneidge, S. A. (1985) EMBO J. 4, 1471-1477 3. Okada, M., Howell, B. W., Broome, M. A., and Cooper, J. A. (1993) J. Biol. Chem. 268, 18070-18075 4. Leiming Li, Masaya Okura, Akira Imamoto MOLECULAR AND CELLULAR BIOLOGY, Feb. 2002, p. 1203
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