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Technical Data Sheet

AM1141

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Akt (Ser-473), phospho-specific
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$295
200 μl
Hu, Rt, Ms
60 kDa

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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).

Application
Dilution


ELISA
1:2000


IP
1:100


WB
1:250



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Wu, C. et al. (2012) Cell. 148(5):973. WB: mouse embryonic fibroblasts
Paterniti, I. et al. (2011) Eur J Neurosci. 33(8):411. WB: mouse spinal cord
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Background
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.


Background References
Jones, P.F. et al. (1991) Proc Natl Acad Sci. USA 88:4171.
Marte, B. & Downward. J. (1997) TIBS. 22:355-358.
Powell, D.J. et al. (2003) Mol. Cell Biol. 23:7794-7808.
Immunogen
Clone M114 was generated from a peptide containing amino acid residues surrounding Serine 473 in human Akt1. This sequence is highly conserved in human and mouse Akt, and may recognize Akt2 and Akt3.
Buffer and Storage
Mouse monoclonal purified with protein G chromatography is supplied in 200μl phosphate-buffered saline with 0.02% sodium azide. Store at 4°C, stable for 6 months. For long term storage, aliquot and store at -20°C.
Specificity
This antibody detects a 60 kDa* protein corresponding to the apparent molecular mass of Akt on SDS-PAGE immunoblots of mouse NIH3T3 cells treated with PDGF and human A431 cells treated with EGF.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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