Western blot using AKT1 polyclonal antibody (Cat # PAB9932) shows detection of endogenous protein in whole cell extracts from H2O2 treated MCF-7 cells (Lane 2). Recombinant AKT1 is detected in lane 3. Lanes 1 and 4 contain wcl from HeLa and A-431 cells, respectively. Specific band staining is completely blocked when antibody is pre-incubated with the immunizing peptide (data not shown). The band at ~55-60 KDa, indicated by the arrowhead, corresponds to the expected molecular weight of AKT1. Primary antibody was diluted 1 : 300 in 1% BLOTTO and reacted with the membrane overnight at 4°C. IRDYE800™ conjugated Dnky-a-Sheep IgG was used at a 1 : 10,000 dilution for 45 min at room temperature.
The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq