SRC Polyclonal Antibody
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SRC Polyclonal Antibody
Background:
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; SRC_HUMAN.Gene ID:
6714Swiss Prot:
P12931Cellular Locus:
Cytoplasm, Nucleus, Cell membraneHost:
RabbitCross Reactivity:
Human, Mouse, RatImmunogen:
2-100/541Target:
SRCClonality:
PolyclonalIsotype:
IgGConjugation:
UnconjugatedSource:
KLH conjugated synthetic peptide derived from human SRCApplications:
WB, ELISA, IHC-P, IHC-F, IF (IHC-P), IF (IHC-F), IF (ICC)Purification:
Purified by Protein A.Concentration:
1µg/µlDilution:
WB (1:300-5000), ELISA (1:500-1000), IHC-P (1:200-400), IHC-F (1:100-500), IF (IHC-P) (1:50-200), IF (IHC-F) (1:50-200), IF (ICC) (1:50-200)Buffer:
0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.Modification:
UnmodifiedStorage Conditions:
Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.Gene ID URL:
6714Predicted Cross Reactivity:
Dog, Cow
