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9272
Akt Antibody
Primary Antibodies
Polyclonal Antibody

Akt Antibody #9272

Citations (10213)
Western blot analysis of extracts from CHO cells, transfected with non-targeted (-) or SignalSilence® Akt siRNA I (+) siRNA, using Akt Antibody #9272 and p42 MAP Kinase (Erk2) Antibody #9108. The Akt antibody confirms silencing of protein expression while the p42 MAP Kinase (Erk2) antibody was used to control for loading and specificity of Akt siRNA (A). Phospho-GSK-3β (Ser9) Antibody #9336 was used to confirm downstream pathway inhibition (B).
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Akt siRNA I (+), using Akt Antibody #9272 and p42 MAP Kinase (Erk2) Antibody #9108. Akt antibody confirms silencing of Akt expression, while the p42 MAP kinase (Erk2) antibody is used to control for loading and specificity of Akt siRNA.
Western blot analysis of extracts from NIH/3T3 cells, untreated or PDGF-treated (50 ng/ml) for the indicated times, using Phospho-Akt (Ser473) Antibody #9271 (upper) or Akt Antibody (lower).
Immunoprecipitation of Akt from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Akt Antibody. Western blot was performed using Akt Antibody. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.
Confocal immunofluorescent images of C2C12 cells showing nuclear and cytoplasmic localization with Akt Antibody (left, red) compared to an isotype control (right). Actin filaments have been labeled with fluorescein phalloidin.
Flow cytometric analysis of Jurkat cells using Akt Antibody (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:200
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Akt Antibody detects endogenous levels of total Akt1, Akt2 and Akt3 proteins. The antibody does not cross-react with related kinases.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey, Chicken, D. melanogaster, Bovine, Dog, Pig, Guinea Pig

Species predicted to react based on 100% sequence homology:

Dog

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy-terminal sequence of mouse Akt. 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 cell 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 PI3K/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).

  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.

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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To Purchase # 9272
Cat. # Size Qty. Price
9272S
100 µl