CST, 9271L, Phospho-Akt (Ser473) Antibody

Polyclonal Antibody for studying Akt2 (Ser474) phosphate/Akt3 (Ser472) phosphate/Akt1 (Ser473) phosphate. Validated for WB,IP,IF,F. Available in 3 sizes. Highly specific and rigorously validated in-house, Phospho-Akt (Ser473) Antibody (CST #9271) is ready to ship. Product Usage Information Western Blotting: 1:1000 Immunoprecipitation: 1:100 Immunofluorescence (Immunocytochemistry): 1:50 - 1:200 Flow Cytometry (Fixed/Permeabilized): 1:100 - 1:400 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized) Specificity / Sensitivity Phospho-Akt (Ser473) Antibody detects endogenous levels of Akt1 only when phosphorylated at Ser473. This antibody also recognizes Akt2 and Akt3 when phosphorylated at the corresponding residues. This antibody does not recognize Akt phosphorylated at other sites, nor does it recognize phosphorylated forms of related kinases such as PKC or p70 S6 kinase. Species Reactivity: Human, Mouse, Rat, Hamster, Monkey, D. melanogaster, Bovine, Dog Source / Purification Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser473 of mouse Akt. Antibodies are purified by protein A and peptide affinity chromatography. Background Akt, also referred to as PKB or Rac, plays a critical role in controlling cell survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3K/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin-dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19). Alternate Names AKT; akt 2; AKT serine/threonine kinase 1; AKT serine/threonine kinase 2; AKT serine/threonine kinase 3; AKT1; AKT1 kinase; AKT1m; AKT2; AKT3; DKFZp434N0250; HIHGHH; MGC99656; MPPH; MPPH2; Murine thymoma viral (v-akt) homolog-2; PKB; PKB alpha; PKB beta; PKB gamma; PKB-ALPHA; PKB-GAMMA; PKBB; PKBBETA; PKBG; PRKBA; PRKBB; PRKBG; Protein kinase Akt-2; Protein kinase Akt-3; Protein kinase B; Protein kinase B alpha; Protein kinase B beta; Protein kinase B gamma; Proto-oncogene c-Akt; putative v-akt murine thymoma viral oncoprotein 2; RAC; rac protein kinase alpha; rac protein kinase beta; RAC-ALPHA; RAC-alpha serine/threonine-protein kinase; RAC-BETA; RAC-beta serine/threonine-protein kinase; RAC-gamma; RAC-gamma serine/threonine protein kinase; RAC-gamma serine/threonine-protein kinase; RAC-PK-alpha; RAC-PK-beta; RAC-PK-gamma; serine threonine protein kinase, Akt-3; serine-threonine protein kinase; STK-2; v-akt murine thymoma viral oncogene homolog 1; v-akt murine thymoma viral oncogene homolog 2; v-akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma); v-akt murine thymoma viral oncogene-like protein 1 Specification REACTIVITY: H M R Hm Mk Dm B Dg SENSITIVITY: Endogenous MW (kDa): 60 SOURCE: Rabbit