Product Pathways - Akt Signaling

Phospho-Akt Pathway Sampler Kit #9916

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Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Source
Phospho-Akt (Ser473) (D9E) Rabbit mAb # 4060	40 microliters	W IP IHC-P IF-IC F	H M R Dr Z	60	Rabbit
Phospho-Akt (Thr308) (C31E5E) Rabbit mAb # 2965	40 microliters	W IF-IC F	H M R Mk	60	Rabbit
Akt (pan) (C67E7) Rabbit mAb # 4691	40 microliters	W IP IHC-P IF-IC F	H M R Mk	60	Rabbit
Phospho-c-Raf (Ser259) Antibody # 9421	40 microliters	W IP	H M R X (C)	74	Rabbit
Phospho-GSK-3β (Ser9) Antibody # 9336	40 microliters	W	H M R Mk (Z)	46	Rabbit
Phospho-PTEN (Ser380) Antibody # 9551	40 microliters	W IP	H M R	54	Rabbit
Phospho-PDK1 (Ser241) Antibody # 3061	40 microliters	W IP	H M R	58 to 68	Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074	100 microliters				Goat
LY294002 (PI3 Kinase Inhibitor) # 9901	0.3 milligrams			307

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken  Dr=Drosophila  X=Xenopus  Z=Zebra Fish

Specificity / Sensitivity

Each phospho-specific antibody recognizes the phosphorylated form of its target. Akt Antibody recognizes total Akt protein, independent of its phosphorylation state.

Source / Purification

Antibodies are produced by immunizing rabbits with synthetic phospho-peptides corresponding to residues surrounding Ser473 or Thr308 of mouse Akt, Ser9 of human GSK-3beta, Ser259 of human c-Raf, Ser380 of human PTEN or Ser241 of human PDK1. Polyclonal 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 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 by phosphorylating and inactivating 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). 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 (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. Nave, 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.

Application References

• Crowder, R.J. and Freeman, R.S. (2000) J Biol Chem 275, 34266-71. This article references the use of Phospho-Akt Pathway Sampler Kit in the following applications: Western Blotting
• Guo, M. et al. (2000) Biochem Pharmacol 60, 635-42. This article references the use of Phospho-Akt Pathway Sampler Kit in the following applications: PI3K inhibition

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