Catalog # AP1001

Akt (Thr-34), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Rb

MW 60 kDa

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 (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).

Phospho-Akt (Thr-34) synthetic peptide (coupled to KLH) corresponding to amino acid residues around threonine 34 of human Akt. This sequence is highly conserved in rat and mouse Akt.

*For more information, see UniProt Accession P31749

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium 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.

The antibody detects a 60 kDa* protein corresponding to the apparent molecular mass of phoshorylated Akt on SDS-PAGE immunoblots of A431 + calyculin A cell lysate. Similar results were seen in calyculin A treated human aortic endothelial and HeLa cells, rabbit spleen fibroblasts, and rat pituitary cells.

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

This kit contains: