Product Pathways - Akt Signaling
Akt1 (2H10) Mouse mAb #2967
| Applications | Reactivity | MW (kDa) | Source | Isotype |
|---|---|---|---|---|
| W IP IF-IC F | H M R | 60 | Mouse | IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
The Akt1 isoform specific (2H10) Mouse mAb detects endogenous levels of Akt1, regardless of its phosphorylation state and does not cross-react with Akt 2 and Akt 3.
Source / Purification
Monoclonal antibody is produced by immunizing mice with a synthetic peptide (KLH-coupled). Antibody is purified by protein G affinity chromatography.
Western Blotting
Western blot analysis of Akt1 (lane1), Akt2 (lane2) and Akt3 (lane3) recombinant proteins, and extracts of NIH/3T3, HeLa, and COS cells, using Akt1 (2H10) Mouse mAb.
Flow Cytometry
Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or PDGF-treated (green), using Akt1 (2H10) Mouse mAb compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent analysis of C2C12 cells showing nuclear and cytoplasmic localization with Akt1 (2H10) Mouse mAb (left, red) compared to an isotype control (right). Actin filaments have been labeled with fluorescein phalloidin.
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 by phosphorylating and inactivating 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). Inhibition of mTOR stops the protein synthesis machinery due to inactivation of its effector, p70 S6 kinase and activation of the eukaryotic initiation factor 4E binding protein 1 (4E-EP1), an inhibitor of translation (18,19).
Three mammalian Akt isoforms have been identified: Akt1, Akt2, and Akt3. Akt2 and Akt3 have 81% and 83% identity in amino acid sequences with Akt1, respectively (18).
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Application References
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