Product Pathways - PI3K / Akt Signaling
PhosphoPlus® Akt (Ser473) Antibody Kit #9270
|9270S||1 Kit (10 western blots)||---||In Stock||---|
|9270||carrier free and custom formulation / quantity||email request|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||Mol. Wt.||Isotype|
|Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060||100 µl||W, IP, IHC-P, IHC-F, IF-IC, F||H, M, R, Hm, Mk, Dm, Z, B||C, X, Dg, Pg||60 kDa||Rabbit IgG|
|Akt (pan) (C67E7) Rabbit mAb #4691||100 µl||W, IP, IHC-P, IF-IC, F||H, M, R, Mk, Dm||Pg||60 kDa||Rabbit IgG|
|Akt Control Cell Extracts #9273||200 µl|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-biotin, HRP-linked Antibody #7075||100 µl||Goat|
|Biotinylated Protein Ladder Detection Pack #7727||100 µl|
|20X LumiGLO® Reagent and 20X Peroxide #7003||5 ml each||W|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey, Dm=D. melanogaster, Z=Zebrafish, B=Bovine
Western blot analysis of extracts from PC3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum- starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060.
Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb #4691.
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002, wortmannin and U0126 (blue), using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060 compared to a nonspecific negative control antibody (red).
The PhosphoPlus® Akt (Ser473) Antibody Kit provides reagents and protocols for the rapid analysis of Akt phosphorylation.
Specificity / Sensitivity
Phospho-Akt (Ser473) Antibody detects endogenous levels of Akt1 only when phosphorylated at Ser473, and Akt2 and Akt3 when phosphorylated at equivalent sites. 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. Akt Antibody detects endogenous levels of total Akt1, Akt2 and Akt3 proteins. It does not cross-react with related kinases.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser473 or the carboxy-terminal sequence of Akt.
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 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).
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- Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
- Franke, T.F. et al. (1995) Cell 81, 727-36.
- 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.
- Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.
- Cross, D.A. et al. (1995) Nature 378, 785-9.
- Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.
- 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.
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* Product-specific protocol.
- 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
- 5106 Phospho-Akt (Thr308) (L32A4) Mouse mAb
- 4051 Phospho-Akt (Ser473) (587F11) Mouse mAb
- 3787 Phospho-Akt (Ser473) (736E11) Rabbit mAb
- 2938 Akt1 (C73H10) Rabbit mAb
- 2967 Akt1 (2H10) Mouse mAb
- 2964 Akt2 (5B5) 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
For Research Use Only. Not For Use In Diagnostic Procedures.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.