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Western blot analysis of Akt (pT308). Lysates from control (left panel) and PDGF-treated (right panel) NIH/3T3 mouse embryonic fibroblasts were probed with purified mAb J1-223.371 at concentrations of 4.0 (lanes 1 and 4), 2.0 (lanes 2 and 5), and 1.0 (lanes 3 and 6) µg/ml. Akt (pT308) is identified as a band of 60 kDa in the treated lysate.
Western blot analysis of Akt (pT308). Lysates from control (left panel) and PDGF-treated (right panel) NIH/3T3 mouse embryonic fibroblasts were probed with purified mAb J1-223.371 at concentrations of 4.0 (lanes 1 and 4), 2.0 (lanes 2 and 5), and 1.0 (lanes 3 and 6) µg/ml. Akt (pT308) is identified as a band of 60 kDa in the treated lysate.
Western blot analysis of Akt (pT308). Lysates from control (left panel) and PDGF-treated (right panel) NIH/3T3 mouse embryonic fibroblasts were probed with purified mAb J1-223.371 at concentrations of 4.0 (lanes 1 and 4), 2.0 (lanes 2 and 5), and 1.0 (lanes 3 and 6) µg/ml. Akt (pT308) is identified as a band of 60 kDa in the treated lysate.
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Akt [also known as PKB (Protein kinase B) or RAC-PK (Related to the A and C kinases)] is a family of serine/threonine kinases that contains a pleckstrin homology (PH) domain. PH domains play important roles in signal transduction. There are three known isoforms of Akt in mammalian cells [Akt1 (α), Akt2 (β) and Akt3 (γ)]; they are thought to be regulated similarly. Akt is activated by insulin and growth factors by a mechanism involving phosphoinositide 3-OH kinase. Phosphoinositide 3-OH kinase products bind to the PH domain, resulting in translocation of Akt to the plasma membrane and activation of Akt to phospho-Akt by upstream kinases. Akt is phosphorylated within the activation loop at threonine 308 (T308) and the C-terminus at serine 473. Phospho-Akt promotes cell survival by inhibiting apoptosis. Specifically, phospho-Akt1 has been shown to phosphorylate Bad, a member of the Bcl-2 family that promotes cell death. This phosphorylation results in the inactivation of the proapoptotic function of Bad. The Akt molecule is thus considered to link extracellular survival signals (growth factors) with the apoptotic machinery (Bad). Akt is also a key mediator of the metabolic effects of insulin. Additionally, Akt has been referred to as an oncogene because it has increased activity in a number of tumors.
The J1-223.371 antibody recognizes Akt phosphorylated at T308.
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