Product Pathways - PI3K / Akt Signaling
Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) #8599
|W IP||H M R||Endogenous||60||Rabbit IgG|
Reactivity Key: H=Human M=Mouse R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) recognizes endogenous levels of Akt2 protein only when phosphorylated at Ser474. This antibody does not cross-react with Akt1 protein when phosphorylated at Ser473 or with Akt3 protein when phosphorylated at Ser472.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser474 of human Akt2 protein.
Western blot analysis of extracts from Akt1 (-/-) mouse embryonic fibroblasts (MEF) or Akt2 (-/-) MEF, untreated (-) or treated with Human Platelet-Derived Growth Factor AA (hPDGF-AA) #8913 (100 ng/ml, 15 min; +), using Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle), or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Western blot analysis of extracts from LNCaP cells, transfected with a construct expressing Akt1 shRNA or Akt2 shRNA, uninduced (-) or doxycycline-induced for the indicated times, using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) #9018 (top), Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) (2nd from top), Akt1 (C73H10) Rabbit mAb #2938 (middle), Akt2 (D6G4) Rabbit mAb #3063 (2nd from bottom), or PI3 Kinase p85 (19H8) Rabbit mAb #4257 (bottom). (Data kindly provided by Drs. Rebecca Chin and Alex Toker, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA).
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).
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- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
- 9998 BSA
- 9999 Nonfat Dry Milk
For Research Use Only. Not For Use In Diagnostic Procedures.