YOUR CART
c-Src (proto-oncogene tyrosine-protein kinase Src) is a 60 kDa tyrosine protein kinase that contains both SH2 and SH3 domains. c-Src is a lipid-modified signaling molecule (myristoylated) involved in cell-cell interactions, cell migration and proliferation through the phosphorylation of many substrates. c-Src is expressed at high levels in differentiated cells including neurons, platelets and macrophages, and can be activated by phospholipase D, insulin-like growth factor receptor, EGF-receptor, fibroblast growth factor receptor and prolactin receptor binding. c-Src can be phosphorylated on multiple sites to interact with polyoma virus middle T antigen, PYK2, HSP72 and caveolin, and efficient study methods are in high demand.
The FACE™ Method
In FACE, cells are cultured in 96-well plates and stimulated to induce the pathway of interest. Following stimulation, the cells are fixed rapidly, which preserves activation-specific protein modifications. Each well is then incubated with a primary antibody specific for the activated protein of interest. Subsequent incubation with secondary HRP-conjugated antibody and developing solution provides a colorimetric or chemiluminescent readout that is quantitative and reproducible (Figure 1). The number of cells in each well can be normalized easily with the provided Crystal Violet solution. FACE Kits also contain primary antibody specific for the native inactive protein, so you can monitor both native and activated protein levels in the same experiment. FACE eliminates cellular extractions, radioactive kinase assays, time-consuming Westerns and inefficient epitope interactions that occur on membranes. FACE is a highly sensitive high-throughput assay designed for detecting activated proteins within mammalian cells.
