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PDP - Template Name: Assay Kit Non Radioactive
PDP - Template ID: *******977a420
  1. Products /
  2. Cellular Assay Kits /
  3. Akt Kinase Assay Kit (Nonradioactive)

Akt Kinase Assay Kit (Nonradioactive) #9840

Inquiry Info. # 9840

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    Product Information

    Protocol

    Product Description

    Nonradioactive Akt Kinase Assay Kit provides all the reagents necessary to measure Akt kinase activity in the cell. Immobilized Akt (1G1) mAb is used to immunoprecipitate Akt from cell extracts. Then, an in vitro kinase assay is performed using GSK-3 Fusion Protein as a substrate. Phosphorylation of GSK-3 is measured by Western blotting, using Phospho-GSK-3alpha/beta (Ser21/9) Antibody.

    Specificity / Sensitivity


    Species Reactivity:

    Human, Mouse, Rat

    Background

    Akt, also referred to as PKB or Rac, plays a critical role in controlling cell 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 PI3K/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.

    Database Links

    UniProt ID: P31751 , Q9Y243 , P31749


    Entrez-Gene Id: 208 , 10000 , 207


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