Anti-phospho-Akt (Ser473) (Rac PKa, PKBa)

Akt, also referred to as PKB or Rac, plays a critical role in controlling the balance between survival and apoptosis. This protein kinase is activated by insulin and various growth and survival factors, and functions in a wortmannin-sensitive pathway involving PI3 kinase. Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1, and by phosphorylation within the carboxyterminus at Ser473. Akt promotes cell survival by inhibiting apoptosis through its ability to phosphorylate and inactivate several targets, including Bad, Forkhead transcription factors and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway. LY294002 is a specific PI3 kinase inhibitor. One of the essential functions of Akt is the regulation of glycogen synthesis through phosphorylation and inactivation of glycogen synthase kinase-3a and b. Akt may also play a role in insulin stimulation of glucose transport. In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK3b-mediated phosphorylation and degradation of cyclin D1, and by negatively regulating the cyclindependent kinase inhibitors p27 KIP and p21 WAF1. Akt also plays a critical role in cell growth by directly phosphorylating the mammalian target of rapamycin, mTOR, but more importantly through phosphorylation and inactivation of tuberin (TSC2), an mTOR inhibitor. Inhibition of mTOR stops the protein synthesis machinery due to inactivation of its effector, p70 S6 kinase, and activation of the eukaryotic initiation factor, 4E binding protein 1 (4E-EP1), an inhibitor of translation. Applications: Suitable for use in Flow Cytometry, Western Blot, Immunofluorescence and Immunoprecipitation. Other applications not tested. Recommended Dilution: Western Blot: 1:1000, Immunoprecipitation: 1:100, Flow Cytometry: 1:25, Immunofluorescence (IC): 1:25, Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.