Technical Data
A1124-02Q
Akt, phosphorylated (Ser473) (AKT1, Proto-oncogene c-Akt, MGC99656, Protein Kinase B, PKB, PKB-alpha, PRKBA, RAC, RAC-alpha Serine/threonine-protein Kinase, RAC-PK-alpha) (PE)
Description:
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-3a and b (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-3b 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 by inactivating p70 S6 kinase and activating the eukaryotic initiation factor 4E binding protein 1 (4E-BP1), an inhibitor of translation (18,19).

Applications:
Suitable for use in Flow Cytometry. Other applications not tested.

Recommended Dilution:
Flow Cytometry: 1:50
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C before opening. DO NOT FREEZE! Stable at 4C as an undiluted liquid. Dilute only prior to immediate use. Stable for at least 12 months at 4C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.
TypeIsotypeCloneGrade
MabIgG11C1015Purified
SizeStorageShippingSourceHost
100ul4CBlue IceHumanRabbit
Concentration:
Not determined
Immunogen:
Synthetic phosphopeptide corresponding to residues surrounding Ser473 of human Akt.
Purity:
Purified
Form
Supplied as a liquid in PBS, pH 7.2, less than 0.1% sodium azide, 2mg/ml BSA. Labeled with R-Phycoerythrin (PE).
Specificity:
Recognizes human Akt, phosphorylated at (Ser473). Species Crossreactivity: mouse, rat, hamster, monkey, mink, chicken, D. melanogaster, xenopus, zebrafish, bovine, canine, porcine, S. cerevisiae, Ce C. elegans, horse.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
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) Nav, 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.