Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Akt Isoform Antibody Sampler Kit #9940

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Akt1 (C73H10) Rabbit mAb #2938 40 µl W IP IHC-P H M R Mk 60 Rabbit IgG
Akt2 (D6G4) Rabbit mAb #3063 40 µl W IP H M R Mk 60 Rabbit IgG
Akt3 (L47B1) Mouse mAb #8018 40 µl W H M R Hm 60 Mouse IgG1
Akt (pan) (C67E7) Rabbit mAb #4691 40 µl W IP IHC-P IF-IC F H M R Mk Dm (Pg) 60 Rabbit IgG
C2C12 cell extracts - Untreated 100 µl
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

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  Hm=Hamster  Mk=Monkey  Dm=D. melanogaster  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each isoform-specific antibody recognizes endogenous levels of the specified Akt isoform independent of its phosphorylation state. Neither antibody cross-reacts with the two other isoforms. Akt Antibody recognizes endogenous levels of all three Akt isoforms.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Akt1 (C73H10) Rabbit mAb #2938.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Akt2 (D6G4) Rabbit mAb #3063 (upper) and Akt1 (C73H10) Rabbit mAb #2938 (lower).

Western Blotting

Western Blotting

Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb #4691.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Akt3 (L47B1) Mouse mAb #8018 (upper) and Akt (pan) (40D4) Mouse mAb #2920 (lower).

Description

Akt Isoform Specific Antibody Sampler Kit provides an economical means of distinguishing between the three Akt isoforms, Akt1, Akt2 and Akt3, as well as a pan-Akt antibody. The kit provides enough primary and secondary antibodies to perform four Western blot experiments.

Source / Purification

Each isoform-specific antibody is produced by immunizing mice or rabbits with a synthetic peptide that is specific for the Akt isoform. Akt1 Monoclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Leu110 of human Akt1. Akt2 monoclonal antibody #3063 is produced by immunizing rabbits with a synthetic peptide corresponding to residues of human Akt2. Akt3 monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg116 of human Akt3 protein. Akt(pan) monoclonal antibody is produced by immunizing animals with a synthetic peptide at the carboxy-terminal sequence of mouse Akt.

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 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.

Application References

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Protocols

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and/or U.S.S.N. 11/476,277) from Epitomics, Inc.

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

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