Akt (Thr-34), Phosphospecific Antibody

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.

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.

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.