DATASHEET
Host:
Rabbit
Target Protein:
SRC
Immunogen Range:
2-100/541
Clonality:
Polyclonal
Isotype:
IgG
Entrez Gene:
6714
Swiss Prot:
P12931
Source:
KLH conjugated synthetic peptide derived from human SRC
Purification:
Purified by Protein A.
Storage Buffer:
0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage:
Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
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.
PRODUCT SPECIFIC PUBLICATIONS
- Jian-xia Wen. et al. Therapeutic Effects and Potential Mechanism of Dehydroevodiamine on N-Methyl-N-Nitro-N-Nitrosoguanidine-Induced Chronic Atrophic Gastritis. Phytomedicine. 2021 Jun;:153619Read more>>
- Huijuan Shen. et al. microRNA-146a mediates distraction osteogenesis via bone mesenchymal stem cell inflammatory response. ACTA HISTOCHEM. 2022 Aug;124:151913Read more>>
- Qi-Lin Lu. et al. Macrophage migration inhibitory factor takes part in the lumbar ligamentum flavum hypertrophy. MOL MED REP. 2022 Sep;26(3):1-9Read more>>
- Hanh Hong Chu. et al. CCL4 Regulates Eosinophil Activation in Eosinophilic Airway Inflammation. INT J MOL SCI. 2022 Jan;23(24):16149Read more>>
- Wei Jiao. et al. Anti-proliferation and anti-migration effect of Yishen Tongbi decoction in experimental rheumatoid arthritis by suppressing SLC3A2/integrin 3 signaling pathways.. PHYTOMEDICINE. 2023 Mar;:154741Read more>>
- Yingmin Liu. et al. LOXL2 promotes tumor proliferation and metastasis by FAK/Src signaling in esophageal squamous cell carcinoma. ELECTRON J BIOTECHN. 2023 ApRead more>>
VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37°C for 20min; Antibody incubation with SRC Polyclonal Antibody, Unconjugated (bs-1135R) at 1:400 overnight at 4°C, followed by a conjugated secondary and DAB staining.
Paraformaldehyde-fixed, paraffin embedded mouse testis; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37°C for 20min; Antibody incubation with SRC Polyclonal Antibody, Unconjugated (bs-1135R) at 1:400 overnight at 4°C, followed by a conjugated secondary and DAB staining.