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.
The mouse fibroblast cell line, SYF, is deficient for the c-Src family kinases, c-Src, Yes, and Fyn, while SYF c-Src overexpress c-Src kinase. This cell line is useful for studies of c-Src mediated phosphorylation when treated with the tyrosine phosphatase inhibitor, pervanadate. Confluent cultures of SYF c-Src cells were serum starved 2 hrs, and then either left untreated (Cat.# SL7171) or treated with pervanadate (1 mM) for 30 minutes at 37°C (cat.# SL7181). Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 10 mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.