YOUR CART
The Janus family of tyrosine kinases (JAK) includes JAK1, JAK2, JAK3 and Tyk2. This family associates with a variety of cytokine receptors and is activated by ligand binding. Upon ligand binding to cytokine receptors, JAKs phosphorylate themselves and their associated receptors. This provides multiple binding sites for signaling proteins containing SH2 or other phospho-tyrosine-binding domains, including STATs, Shc, insulin receptor substrates and FAK. Thus, JAK kinases provide important links between cytokine/hormone receptors and downstream effector proteins, 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.
