anti-Human SRC antibody for Immunofluorescence (Paraffin-embedded Sections)

c-Src tyrosine kinase plays a critical role in signal transduction downstream of growth factor receptors, integrins and G protein-coupled receptors. We used stable isotope labeling with amino acids in cell culture (SILAC) approach to identify additional substrates of c-Src tyrosine kinase in human embryonic kidney 293T cells. We have identified 10 known substrates and interactors of c-Src and Src family kinases along with 26 novel substrates. We have experimentally validated 4 of the novel proteins (NICE-4, RNA binding motif 10, FUSE-binding protein 1 and TRK-fused gene) as direct substrates of c-Src using in vitro kinase assays and cotransfection experiments. Significantly, using a c-Src specific inhibitor, we were also able to implicate 3 novel substrates (RNA binding motif 10, EWS1 and Bcl-2 associated transcription factor) in PDGF signaling. Finally, to identify the exact tyrosine residues that are phosphorylated by c-Src on the novel c-Src substrates, we designed custom peptide microarrays containing all possible tyrosine-containing peptides (312 unique peptides) and their mutant counterparts containing a Tyr --> Phe substitution from 14 of the identified substrates. Using this platform, we identified 34 peptides that are phosphorylated by c-Src. We have demonstrated that SILAC-based quantitative proteomics approach is suitable for identification of substrates of nonreceptor tyrosine kinases and can be coupled with peptide microarrays for high-throughput identification of substrate phosphopeptides.

Synonyms: ASV, Avian sarcoma virus, c SRC, CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA, cSrc, EC 2.7.10.2, Neuronal CSRC tyrosine specic protein kinase, Neuronal SRC, Oncogene SRC, OTTHUMP00000030931, OTTHUMP00000174476, OTTHUMP00000174477, p60 Src, p60-Src, p60Src, pp60c src, pp60c-src, pp60csrc, Proto oncogene tyrosine protein kinase Src, Proto-oncogene c-Src, Proto-oncogene tyrosine-protein kinase Src, Protooncogene SRC, protooncogene SRC Rous sarcoma, Src, SRC Oncogene, SRC_MOUSE, SRC1, Tyrosine kinase pp60c src, Tyrosine protein kinase SRC 1, Tyrosine protein kinase SRC1, v src avian sarcoma Schmidt Ruppin A2 viral oncogene homolog, V src sarcoma Schmidt Ruppin A 2 viral oncogene homolog avian, v src sarcoma Schmidt Ruppin A 2 viral oncogene homolog avian, vsrc avian sarcoma Schmidt Ruppin A2 viral oncogene homolog.