Catalog # SK6350

c-Src Phospho-Regulation Antibody Kit

Antibody Sampler Kit



c-Src was the first proto-oncogenic non-receptor tyrosine kinase characterized in human. The Src family is composed of nine members in vertebrates, including c-Src, Yes, Fgr, Yrk, Fyn, Lyn, Hck, Lck, and Blk. Src-family kinases transduce signals that are involved in the control of a variety of cellular processes, including proliferation, differentiation, motility, and adhesion. Src-family kinases contain an N-terminal cell membrane anchor followed by SH3 and SH2 domains. The activity of c-Src is regulated by tyrosine phosphorylation at multiple sites. Tyrosine 418 is autophosphorylated following c-Src activation. Tyrosine 215 in the SH2 domain of c-Src is phosphorylated following growth factor receptor activation. Both Tyr-215 and Tyr-418 phosphorylation increases tyrosine kinase activity, while phosphorylation of Tyr-530 downregulates c-Src kinase activity. Thus, tyrosine phosphorylation of c-Src is critical for regulating its kinase activity.

Stover, D.R. et al. (1996) J Biol Chem 271(21):12481.
Obergfell, A. et al. (2002) J Cell Biol. 157(2):265.
Vadlamudi, R.K. et al. (2003) FEBS Letters 543:76.

Western blot analysis of mouse SYF cells transformed with c-Src then left untreated (lanes 1, 3, & 5) or treated with pervanadate (1 mM) for 30 minutes (lanes 2, 4, & 6). The blot was probed with anti-c-Src (Tyr-215) (lanes 1 & 2), anti-c-Src (N-terminal region) (lanes 3 & 4), and anti-c-Src (Tyr-530) (lanes 5 & 6).

Western blot analysis of A431 cells treated with pervanadate (1 mM) for 30 minutes (20 µg/lane). The Blot was probed with anti-c-Src (Tyr-215) in the presence of no blocking peptide (lane 1), phospho-c-Src (Tyr-215) peptide (lane 2), unrelated phospho-β-Catenin (Tyr-142) peptide (lane 3), or BSA conjugated to phospho-tyrosine (lane 4). Peptides and BSA were used at 1 µg/ml.

Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
SM2591 Leoni, G. et al. (2013) J Clin Invest. 123(1):443. WB: human intestinal epithelial cells
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

c-Src (N-terminal region) Mouse mAb50 μlWB, E, IPHu, Rt, Ms60
c-Src (Tyr-215)[conserved site], phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms60
c-Src (Tyr-530)[conserved site], phospho-specific Mouse mAb50 μlWB, EHu, Rt, Ms60
Anti-Mouse Ig:HRP Donkey pAb 100 μlWB, EMs
Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb 100 μlWB, E, ICC, IHCRb

The c-Src phospho-regulation antibody sampler kit can be used to detect c-Src phosphorylation on Tyr-215 and Tyr-530. The kit also includes a monoclonal antibody to monitor total c-Src expression levels and secondary reagents for detection of these antibodies.