Phospho-Akt Pathway Antibody Sampler Kit #9916

Kit Usage Information

Protocols

• 3438: Western Blotting, Immunoprecipitation (Magnetic)
• 4060: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
• 4691: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
• 5558: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
• 7074: Western Blotting
• 9421: Western Blotting, Immunoprecipitation (Agarose)
• 9551: Western Blotting, Immunoprecipitation (Magnetic)
• 13038: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Flow

Product Description

The Phospho-Akt Pathway Antibody Sampler Kit provides an economical means to evaluate the activation status of the Akt signaling pathway, including PTEN and phosphorylated Akt, GSK-3beta, c-Raf and PDK1. The kit includes enough primary and secondary antibodies to perform two Western blot experiments.

Specificity / Sensitivity

Each phospho-specific antibody recognizes the phosphorylated form of its target. Akt Antibody recognizes total Akt protein, independent of its phosphorylation state.

Source / Purification

Antibodies are produced by immunizing rabbits with synthetic phosphopeptides corresponding to residues surrounding Ser473 or Thr308 of human Akt, Ser9 of human GSK-3ß, Ser259 of human c-Raf, Ser380 of human PTEN or Ser241 of human PDK1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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 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).

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Select Your Pathway/Protein AMPK Signaling Adherens Junction Dynamics Alzheimer's Disease Angiogenesis Apoptosis Regulation B Cell Receptor Signaling ESC Pluripotency and Differentiation ErbB/HER Signaling G1/S Checkpoint GPCR Signaling to MAPKs Growth And Differentiation Control by MAPKs Hedgehog Signaling IL6 Signaling Inhibition of Apoptosis Insulin Receptor Signaling Microtubule Dynamics Mitochondrial Control of Apoptosis NF-kB Signaling PI3K/Akt Signaling Protein Kinase C Signaling T Cell Receptor Signaling TGF-ß Signaling Translation: eIF2 Translation: eIF4E and p70S6K Warburg Effect Wnt/ß-Catenin Signaling mTOR Signaling Protein: O15530 Protein: P04049 Protein: P31749 Protein: P31751 Protein: P49841 Protein: P60484 Protein: Q9Y243

Database Links

UniProt ID: O15530 , P60484 , P04049 , P31751 , Q9Y243 , P31749 , P49841

Entrez-Gene Id: 5170 , 5728 , 5894 , 208 , 10000 , 207 , 2932

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