Akt (PKB, Rac kinase) is a 60kDa ser/thr kinase critical for controlling diverse cellular functions, including glucose metabolism, gene transcription, cell proliferation, and apoptosis. Akt phosphorylates a number of substrates including MBP, glycogen synthetase, PKA RII subunit, and histone H1. Akt is activated in response to insulin and growth factors in a PI3-kinase dependent manner. Activation of PI3-Kinase generates phosphatidylinositol 3,4-bisphosphate, which induces membrane translocation of Akt coincident with its phosphorylation at Thr-308 and Ser-473. Upon activation, Akt associates with members of the PKC family of kinases, such as PKCδ and PKCζ. Ceramide-activated PKCζ leads to phosphorylation of Thr-34 within the pleckstrin homology domain of Akt. This phosphorylation inhibits PIP3 binding to Akt preventing activation of the kinase and may lead to cermide-induced cell death.
References
Powell, D.J. et al. (2003) Mol. Cell Biol. 23:7794-7808. Marte, B. & Downward. J. (1997) TIBS. 22:355-358.Jones, P.F. et al. (1991) Proc. Natl. Acad. Sci. USA 88:4171-4175.
Western blot analysis of A431 cells, serum starved overnight (lanes 1 & 3) or treated with calyculin A (100 nM) for 30 min. (lanes 2 & 4). The blot was probed with anti-Akt (Thr-34) (lanes 1 & 2) or anti-Akt1 (N-terminal region) (lanes 3 & 4).
Western blot analysis of A431 cells untreated (lanes 1 & 3) or treated with 100 ng/ml EGF for 60 min. (lanes 2 & 4). The blots were probed with monoclonal anti-phospho-Akt (Ser-473) (lanes 1 & 2) and monoclonal anti-Akt1 (N-terminal region) (lanes 3 &4).
*For more information, see UniProt Accession P31749
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*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blotmobilities of known proteins with similar MW.
Product References:
Wei, H. & Vander Heide, R. (2010) AJP Heart Circ. 298:H152. (WB: rat myocardium)Li, W. et al. (2009) Am J Physiol Cell Physiol (2009) 297:706. (WB: mouse C2C12 cells)Chambers, M.A. et al. (2009) J Physiol. 587:3363. (WB: mouse muscle, C2C12)Moylan, J.S. et al. (2008) AJP Cell. Phys. 295:986. (WB: mouse C2C12)Smith, M. et al. (2007) Am J Physiol Cell Physiol 293:1947. (WB: mouse C2C12 cells)This kit contains:
