Akt3 Antibody

4059:

Immunoprecipitation , Western Blotting

Akt3 Antibody detects endogenous levels of total Akt3, but does not recognize the truncated form of rat Akt3. The antibody does not cross-react with recombinant Akt1 or Akt2.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues of human Akt3. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from CAD and PC12 cells, using Akt3 Antibody.

Western Blotting

Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from MDA-MB-231 cells, using Akt3 Antibody (upper) or Akt Antibody #9272 (lower). Recombinant Akt3 is a 6His-fusion protein, MW=66 kDa.

IP

Western blot analysis of immunoprecipitates from MDA-MB-231 cell lysates. Akt1 was immunoprecipitated with Akt1 (2H10) Monoclonal Antibody #2967, Akt 2 was immunoprecipitated with Akt2 Antibody #2962, Akt 3 was immunoprecipitated with Akt3 Antibody, and specificity was confirmed by immunprecipitation with rabbit IgG. Immunoblot was performed using Akt1 Antibody #2967 (left), Akt2 Antibody #2962 (middle) and Akt3 Antibody #4059 (right).

IP

Western blot analysis (lane 1) and Immunoprecipitation (lanes 2-5) of extracts from HA-Akt3-transfected HeLa cells. HA-Akt3 was immunoprecipitated with HA-Tag (6E2) Mouse mAb #2367 (lane 2) or with Akt3 Antibody (lane 4). Immunprecipitation with mouse IgG (lane 3) and rabbit IgG (lane 5) confirm specificity. Immunoblot was performed using HA-Tag (6E2) Mouse mAb #2367.

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.

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• 3788 Akt3 (62A8) Rabbit mAb
• 4060 Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb
• 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
• 4691 Akt (pan) (C67E7) Rabbit mAb
• 2938 Akt1 (C73H10) Rabbit mAb
• 2967 Akt1 (2H10) Mouse mAb
• 2964 Akt2 (5B5) Rabbit mAb
• 9916 Phospho-Akt Pathway Antibody Sampler Kit
• 9273 Akt Control Cell Extracts
• 9901 LY294002 (PI3 Kinase Inhibitor)
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 7727 Biotinylated Protein Ladder Detection Pack
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)

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