Product Pathways - PI3K / Akt Signaling
PhosphoPlus® Akt Activation Kit #9280
|9280S||1 Kit (10 western blots)||---||In Stock||---|
|9280||carrier free and custom formulation / quantity||email request|
|Kit Includes||Quantity||Applications||Reactivity||Homology†||Mol. Wt.||Isotype|
|Phospho-Akt (Thr308) (C31E5E) Rabbit mAb #2965||100 µl||W, IF-IC, F||H, M, R, Hm, Mk||60 kDa||Rabbit IgG|
|Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060||100 µl||W, IP, IHC-P, IHC-F, IF-IC, F||H, M, R, Hm, Mk, Dm, Z, B||C, X, Dg, Pg||60 kDa||Rabbit IgG|
|Akt (pan) (C67E7) Rabbit mAb #4691||100 µl||W, IP, IHC-P, IF-IC, F||H, M, R, Mk, Dm||Pg||60 kDa||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-biotin, HRP-linked Antibody #7075||100 µl||Goat|
|Biotinylated Protein Ladder Detection Pack #7727||100 µl|
|20X LumiGLO® Reagent and 20X Peroxide #7003||5 ml each||W|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen)
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey, Dm=D. melanogaster, Z=Zebrafish, B=Bovine
Western blot analysis of extracts from NIH/3T3 and Jurkat cells, untreated, PDGF-treated or LY294002-treated as indicated, using Phospho-Akt (Thr308) (C31E5) Rabbit mAb #2965 (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Western blot analysis of extracts from PC3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum- starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060.
Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue) using Phospho-Akt (Thr308) (C31E5) Rabbit mAb #2965.
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002, wortmannin and U0126 (blue), using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060 compared to a nonspecific negative control antibody (red).
7003 5 ml
Specificity / Sensitivity
The phospho-Akt antibodies detect endogenous levels of Akt only when phosphorylated at either Ser473 or Thr308. The Akt antibody detects endogenous levels of total Akt1, Akt2 and Akt3 proteins independent of their phosphorylation state. It does not cross-react with related kinases.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding either Ser473, Thr308 or from the carboxy-terminal sequence of mouse Akt.
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).
The Akt activation kit provides an economical means to evaluate the activation status of Akt at both the Ser473 and Thr308 positions as well as the total Akt protein level. The kit includes enough primary and secondary antibodies to perform ten Western mini-blot experiments.
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- Brunet, A. et al. (1999) Cell 96, 857-68.
- Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
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- Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
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- Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
- Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
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- Manning, B.D. et al. (2002) Mol Cell 10, 151-62.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
* Product-specific protocol.
- 4058 Phospho-Akt (Ser473) (193H12) Rabbit mAb
- 4056 Phospho-Akt (Thr308) (244F9) Rabbit mAb
- 2938 Akt1 (C73H10) Rabbit mAb
- 9916 Phospho-Akt Pathway Antibody Sampler Kit
- 9940 Akt Isoform Antibody Sampler Kit
- 7160 PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Kit
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 3063 Akt2 (D6G4) Rabbit mAb
- 8018 Akt3 (L47B1) Mouse mAb
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.