CycLex AKT/PKB Kinase Assay/Inhibitor Screening Kit

Other - 1) Screening inhibitors or activators of AKT. 2) Detecting the effects of pharmacological agents on AKT activity.

The AKT oncogene was isolated from the directly transforming murine retrovirus AKT8, which was isolated from an AKR mouse thymoma cell line. Staal cloned the human cellular homolog, AKT1 (1). He found a 20-fold amplification of the AKT1 gene in 1 of 5 gastric adenocarcinomas tested. Phosphoinositide 3-kinases (PI3Ks) generate specific inositol lipids that have been implicated in the regulation of cell growth, proliferation, survival, differentiation and cytoskeletal changes. One of the best characterized targets of PI3K lipid products is the protein kinase AKT or protein kinase B (PKB). In quiescent cells, AKT resides in the cytosol in a low-activity conformation. Upon cellular stimulation, AKT is activated through recruitment to cellular membranes by PI3K lipid products and phosphorylation by 3?-phosphoinositide-dependent kinase-1 (PDK1). Mammals have three closely related AKT genes, encoding the isoforms AKT1, AKT2 and. AKT3 AKT2 and. AKT3 show 81 and 83% amino acid identity with AKT1 respectively. All AKT isoforms show a broad tissue distribution and consist of an N-terminal PH domain, a kinase domain and a C-terminal regulatory tail Two specific sites, one in the kinase domain (Thr308 in AKT1) and the other in the C-terminal regulatory region (Ser473 in AKT1), need to be phosphorylated for full activation of these kinases. AKT was among the first proteins known to contain a PH domain, a few years before the function of this domain came to light. The PH domain of AKT specifically binds PI3K lipid products, and a firm link between PI3K and AKT signaling has now been established. AKT is cytosolic in unstimulated cells, and some of it translocates to the plasma membrane upon activation of PI3K, where it becomes activated (2, 3). Active AKT then appears to detach from the plasma membrane and to translocate through the cytosol to the nucleus. The mechanism of this translocation is unclear. AKT1 was found to mediate insulin- and insulin-like growth factor (IGF-1)-induced cellular responses, such as the inhibition of glycogen synthase kinase-3 (4), the stimulation of glucose uptake (5) and the promotion of cell survival by inhibiting apoptosis (6). Overexpression of AKT1 or AKT2 is associated with some human ovarian, pancreatic, and breast carcinomas (7-9).

The CycLex® AKT/PKB kinase Assay/Inhibitor Screening kit is designed to measure the activities of purified AKTs for the rapid and sensitive evaluation of inhibitors or activators. The phospho-serine specific monoclonal antibody used in this assay kit has been demonstrated to recognize the phospho-serine residue in AKTide-2T, which is efficiently phosphorylated by AKTs.

Human: 207 Mouse: 11651

Microplate, 10x Wash Buffer, Kinase Buffer, 20x ATP, HRP-conjugated Detection Antibody, Substrate Reagent, Stop Solution

AKT1