Product Pathways - PI3K / Akt Signaling
Phospho-Akt (Ser473) Antibody #9271
Have you tried your application using our XP® monoclonal antibodies? Try product: 4060
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP IF-IC F | H M R Hm Dm B Dg Pg (Mk) (C) (X) (Hr) | Endogenous | 60 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Hm=Hamster
Mk=Monkey
C=Chicken
Dm=D. melanogaster
X=Xenopus
B=Bovine
Dg=Dog
Pg=Pig
Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
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. 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.
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.
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.)
Background
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).
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- Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
- Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
- Jacinto, E. et al. (2006) Cell 127, 125-37.
- Cardone, M.H. et al. (1998) Science 282, 1318-21.
- Brunet, A. et al. (1999) Cell 96, 857-68.
- Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
- Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
- Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
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- Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.
- Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
- Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
- Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.
- Manning, B.D. et al. (2002) Mol Cell 10, 151-62.
Application References
- 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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Companion Products
- 4060 Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb
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- 3788 Akt3 (62A8) Rabbit mAb
- 7160 PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Kit
- 7170 PathScan® Total Akt1 Sandwich ELISA Kit
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- 9273 Akt Control Cell Extracts
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- 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
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.