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
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).
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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:
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# | DESCRIPTION | SIZE | APPLICATIONS | SPECIES REACTIVITY | MW (kDa) |
AM1011 | Akt (N-terminal region) Mouse mAb | 50 μl | WB, E, ICC | Hu, Rt, Ms | 60 |
AM1141 | Akt (Ser-473), phospho-specific Mouse mAb | 50 μl | WB, E, IP | Hu, Rt, Ms | 60 |
AP1001 | Akt (Thr-34), phospho-specific Rabbit pAb | 50 μl | WB, E | Hu, Rt, Rb | 60 |
MS3001 | Anti-Mouse Ig:HRP Donkey pAb | 100 μl | WB, E | Ms | |
RS3251 | Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb | 100 μl | WB, E, ICC, IHC | Rb |
KIT SUMMARY
The 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.
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