BackgroundImagesKit ComponentsBuffer & StorageReferencesReviews 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. ReferencesJones, P.F. et al. (1991) Proc. Natl. Acad. Sci. USA 88:4171-4175.Marte, B. & Downward. J. (1997) TIBS. 22:355-358.Powell, D.J. et al. (2003) Mol. Cell Biol. 23:7794-7808. 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).Western blot analysis of A431 cells, serum starved overnight (20 µg/lanes 1 & 3) and calyculin A (100 nM) treated for 30 minutes (20 µg/lanes 2 & 4). The blot was probed with anti-Akt (Thr-34) (lanes 1 & 2) or anti-Akt1 (N-terminal region) (lanes 3 & 4). AP1001 and AM1011 are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide, and AM1141 is provided in PBS with 0.02% azide. The secondary reagents are supplied in PBS, 50% glycerol, 1 mg/ml BSA, and no azide. Store at –20°C. Stable for 1 year. The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.*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:RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblastsAM1141 Wu, C. et al. (2012) Cell. 148(5):973. WB: mouse embryonic fibroblastsAM1141 Paterniti, I. et al. (2011) Eur J Neurosci. 33(8):411. WB: mouse spinal cordMS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western BlotRS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western BlotAM1011 Wei, H., Vander Heide, R. (2010) AJP Heart Circ. 298:H152. WB: rat myocardiumAM1011 Chambers, M.A. et al. (2009) J Physiol. 587:3363. WB: mouse muscle, C2C12AM1011 Moylan, J.S. et al. (2008) AJP Cell. Phys. 295:986. WB: mouse C2C12This kit contains:CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)AM1011Akt (N-terminal region) Mouse mAb50 μlWB, E, ICCHu, Rt, Ms60AM1141Akt (Ser-473), phospho-specific Mouse mAb50 μlWB, E, IPHu, Rt, Ms60AP1001 Akt (Thr-34), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Rb60MS3001Anti-Mouse Ig:HRP Donkey pAb100 μlWB, EMsRS3251Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRbKIT SUMMARYThe Akt phospho-regulation antibody sampler kit can be used to detect Akt activation via Ser-473 phosphorylation, as well as Akt inhibition through Thr-34 phosphorylation. The kit also includes a monoclonal antibody to monitor the total Akt expression levels and secondary reagents for detection of these antibodies.