Phospho-Akt (Ser473) Antibody

9271:

Flow , Immunofluorescence , Immunoprecipitation , Western Blotting

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. It does not recognize Akt phosphorylated at other sites, nor does it recognize phosphorylated forms of related kinases such as PKC or p70 S6 kinase.

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.

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF, wortmannin, LY294002, rapamycin or PD98059, using Phospho-Akt (Ser473) Antibody.

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF for the indicated times, using Phospho-Akt (Ser473) Antibody (upper) or Akt Antibody #9272 (lower).

Western Blotting

Western blot analysis of immunoprecipitated Akt from 293 cells transiently transfected with HA-tagged Akt (WT), HA-tagged K179A mutant Akt and HA-tagged K179A/S473A mutant Akt, using Phospho-Akt (Ser473) Antibody (upper), Akt antibody (middle) or HA antibody (lower). Phospho-Akt (Ser473) Antibody does not recognize Akt with an alanine substituion at Ser473. (Polakiewicz, R.D. et al. [1998] J. Biol. Chem. 273, 23534-23541.)

Flow Cytometry

Flow cytometric analysis of LNCaP cells, untreated (green) or LY294002-treated (blue), using Phospho-Akt (Ser473) Antibody compared to a nonspecific negative control antibody (red).

IF-IC

Confocal immunofluorescent images of C2C12 cells serum starved and treated with or without insulin as indicated and labeled with Phospho-Akt (Ser473) Antibody (red). Actin filaments have been labeled with fluorescein phalloidin.

Akt, also referred to as PKB or Rac, plays a critical role in controlling 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 PI3 kinase/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 Kip (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).

1. Franke, T.F. et al. (1997) Cell 88, 435-7.
2. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
3. Franke, T.F. et al. (1995) Cell 81, 727-36.
4. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
5. Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
6. Jacinto, E. et al. (2006) Cell 127, 125-37.
7. Cardone, M.H. et al. (1998) Science 282, 1318-21.
8. Brunet, A. et al. (1999) Cell 96, 857-68.
9. Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
10. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
11. Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
12. Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.
13. Cross, D.A. et al. (1995) Nature 378, 785-9.
14. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.
15. Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.
16. Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
17. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
18. Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.
19. Manning, B.D. et al. (2002) Mol Cell 10, 151-62.

• Baumann, C. A. et al. (2000) CAP defines a second signalling pathway required for insulin-stimulated glucose transport. Nature 407, 202-207. Applications: Western Blot
• Patrucco, E. et al. (2004) PI3Kgamma modulates the cardiac response to chronic pressure overload by distinct kinase-dependent and -independent effects. Cell 118, 375-387. Applications: Western Blotting
• Fukuda, T. et al. (2003) PINCH-1 is an obligate partner of integrin-linked kinase (ILK) functioning in cell shape modulation, motility, and survival. J. Biol. Chem. 278, 51324-51333. Applications: Western Blotting
• Okkenhaug, K. et al. (2002) Impaired B and T cell antigen receptor signaling in p110delta PI 3-kinase mutant mice. Science 297 (5583), 1031-1034. Applications: Western Blotting
• Wettey, F. R. et al. (2002) Controlled Elimination of Clathrin Heavy-Chain Expression in DT40 Lymphocytes. Science 297, 1521-1525. Applications: Western Blotting
• Sutherland, C. L. et al. (2002) UL16-Binding Proteins, Novel MHC Class I-Related Proteins, Bind to NKG2D and Activate Multiple Signaling Pathways in Primary NK Cells. J. Immunol. 168, 671-679. Applications: Western Blotting
• Holland, E. C. et al. (2000) Combined activation of Ras and Akt in neural progenitors induces glioblastoma formation in mice. Nat. Genet. 25, 55-57. Applications: Western Blot
• Shan, X. et al. (2000) Deficiency of PTEN in Jurkat T cells causes constitutive localization of Itk to the plasma membrane and hyperresponsiveness to CD3 stimulation. Mol. Cell. Biol. 20, 6945-6957. Applications: Western Blot
• Sibilia, M. et al. (2000) The EGF receptor provides an essential survival signal for SOS-dependent skin tumor development. Cell 102, 211-220. Applications: Western Blot
• Tilton, B. et al. (2000) Signal transduction by CXC chemokine receptor 4: stromal cell-derived factor 1 stimulates prolonged protein kinase b and extracellular signal-regulated kinase 2 activation in T lymphocytes. J. Exp. Med. 192, 313-324. Applications: Western Blot
• Guo, A. et al. (2008) Proc Natl Acad Sci U S A 105, 692-7. Applications: Western Blotting
• Fukuhara, S. et al. (2008) Nat Cell Biol 10, 513-26. Applications: Western Blotting
• Fujita, H. et al. (2005) Biochem Pharmacol 69, 1773-84. Applications: Western Blotting
• Engelman, J.A. et al. (2008) Nat Med , . Applications: Western Blotting
• Hoshino, Y. et al. (2004) Mol Reprod Dev 69, 77-86. Applications: IF-IC (In Cells)
• Chung, Y.W. et al. (2011) J Biol Chem 286, 29681-90. Applications: Western Blotting
• Fonseca, B.D. et al. (2011) J Biol Chem 286, 27111-22. Applications: Western Blotting

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• 4060 Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb
• 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
• 4691 Akt (pan) (C67E7) Rabbit mAb
• 4051 Phospho-Akt (Ser473) (587F11) Mouse mAb
• 9272 Akt Antibody
• 4058 Phospho-Akt (Ser473) (193H12) Rabbit mAb
• 2938 Akt1 (C73H10) Rabbit mAb
• 3788 Akt3 (62A8) Rabbit mAb
• 7160 PathScan^® Phospho-Akt1 (Ser473) Sandwich ELISA Kit
• 7170 PathScan® Total Akt1 Sandwich ELISA Kit
• 9916 Phospho-Akt Pathway Antibody Sampler Kit
• 9901 LY294002 (PI3 Kinase Inhibitor)
• 9273 Akt Control Cell Extracts
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 3063 Akt2 (D6G4) Rabbit mAb

For Research Use Only. Not For Use In Diagnostic Procedures.