AKT, also known as protein kinase B (PKB) or Related to A and C kinases (RAC), is the cellular homolog of the viral oncogene v-AKT. It was originally cloned based on its homology to protein kinases A and C. AKT is activated in response to insulin, and various growth and survival factors. It is clear that AKT enzyme activity is both necessary and essential for cell survival, and AKT has been shown to be able to induce progression of the cell cycle as well as suppress apoptosis. The key role of AKT in the regulation of cell survival makes it a promising target for anti-oncogenic therapeutics.
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.


