Focal adhesion kinase (FAK) is a widely expressed non-receptor cytoplasmic tyrosine kinase that is implicated in integrin-mediated signal transduction. FAK plays an important role in the control of several biological processes, including cell spreading, migration and survival. Physical interactions of FAK with the integrin cytoplasmic domain and cytoskeletal proteins talin, paxillin and/or tensin play a key role in FAK activation by facilitating its oligomerization and transphosphorylation. FAK shows a rapid increase in tyrosine phosphorylation when stimulated by diverse signaling molecules, including those that regulate embryonic development, cell proliferation, migration and apoptosis, 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.


