Akt (5G3) Mouse mAb (Alexa Flu

Product Pathways - PI3K / Akt Signaling

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Akt (5G3) Mouse mAb (Alexa Fluor^® 488 Conjugate) #2917

Applications	Reactivity	Sensitivity	Isotype
F	H M R Hm	Endogenous	Mouse IgG1

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

2917:

Flow

Specificity / Sensitivity

Akt (5G3) Mouse mAb (Alexa Fluor^® 488 Conjugate) detects endogenous levels of Akt. This antibody does not cross-react with other related proteins.

Source / Purification

Monoclonal antibody is produced by immunizing animals with an Akt1 recombinant protein containing human Akt1 residues 140-480. The antibody was conjugated to Alexa Fluor^® 488 under optimal conditions with an F/P ratio of 2-6.

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor^® 488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #2966 reacts with human, mouse, rat and hamster Akt protein. CST expects that Akt (5G3) Mouse mAb (Alexa Fluor^® 488 Conjugate) will also recognize Akt in these species.

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).

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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !

Companion Products

• 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
• 4075 Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate)
• 4691 Akt (pan) (C67E7) Rabbit mAb
• 9272 Akt Antibody
• 9901 LY294002 (PI3 Kinase Inhibitor)
• 2975 Rabbit (DA1E) mAb IgG XP^® Isotype Control (Alexa Fluor^® 488 Conjugate)
• 2966 Akt (5G3) Mouse mAb
• 2964 Akt2 (5B5) Rabbit mAb
• 3788 Akt3 (62A8) Rabbit mAb

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