Product Pathways - Motif Antibodies

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Phospho-(Ser/Thr) Akt Substrate Antibody #9611

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Applications	Reactivity	Sensitivity	Source
W IP IHC-P E-P	All	Endogenous	Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

9611:

ELISA Peptide, IHC / Paraffin, Immunoprecipitation, Western Blotting

Specificity / Sensitivity

Phospho-(Ser/Thr) Akt Substrate Antibody preferentially recognizes peptides and proteins containing phospho-Ser/Thr preceded by Lys/Arg at positions -5 and -3, in a manner largely independent of other surrounding amino acids. Some cross-reactivity is observed for peptides that contain phospho-Ser/Thr preceded by Arg/Lys at positions -3 and -2. No cross-reactivity is observed with the corresponding nonphosphorylated sequences or with other phospho-Ser/Thr-containing motifs. By ELISA, the antibody recognizes a wide range of phosphorylated Akt substrate peptides, and, by 2-D gel Western blot analysis, it recognizes a large number of proteins presumed to be Akt substrates. (U.S. Patent No's.: 6,441,140; 6,982,318; 7,259,022; 7,344,714; U.S.S.N. 11,484,485; and all foreign equivalents.)

Source / Purification

Polyclonal antibodies are produced by immunizing animals with phospho-Akt substrate peptides . Antibodies are purified by protein A and peptide affinity chromatography.

Background

An important class of kinases, referred to as Arg-directed kinases or AGC-family kinases, includes cAMP-dependent protein kinase (PKA), cGMP-dependent protein kinase (PKG), protein kinase C, Akt, and RSK. These kinases share a substrate specificity characterized by Arg at position -3 relative to the phosphorylated Ser or Thr (1,2). Akt plays a central role in mediating critical cellular responses including cell growth and survival, angiogenesis, and transcriptional regulation (3-5). While a number of Akt substrates are known (such as GSK-3, Bad, and caspase-9) many important substrates await discovery. Akt phosphorylates substrates only at Ser/Thr in a conserved motif characterized by Arg at positions -5 and -3 (6). Phospho-Akt substrate-specific antibodies from Cell Signaling Technology are powerful tools for investigating the regulation of phosphorylation by Akt and other Arg-directed kinases, as well as for high throughput kinase drug discovery.

1. Montminy, M. (1997) Annu Rev Biochem 66, 807-22.
2. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
3. Marte, B.M. and Downward, J. (1997) Trends Biochem Sci 22, 355-8.
4. Jiang, B.H. et al. (2000) Proc Natl Acad Sci USA 97, 1749-53.
5. Scheid, M.P. and Woodgett, J.R. (2000) Curr Biol 10, R191-4.
6. Alessi, D.R. et al. (1996) FEBS Lett 399, 333-8.

Application References

• Fonseca, B.D. et al. (2011) J Biol Chem 286, 27111-22. Applications: Western Blotting
• Højlund, K. et al. (2008) Diabetes 57, 357-66. Applications: IP Western Blotting
• Karlsson, H.K. et al. (2006) Diabetes 55, 1283-8. Applications: Western Blotting
• Srirangam, A. et al. (2006) Clin Cancer Res 12, 1883-96. Applications: IHC-P (paraffin)
• Nascimento, E.B. et al. (2006) Diabetes 55, 3221-8. Applications: 2D Analysis Western Blotting
• Trinidad, A.G. et al. (2006) J Leukoc Biol 79, 1073-82. Applications: Western Blotting
• Zhao, H. et al. (2005) J Neurosci 25, 9794-806. Applications: IF-P (paraffin)
• Bruss, M.D. et al. (2005) Diabetes 54, 41-50. Applications: Western Blotting
• Wada, S. et al. (2005) J Biol Chem 280, 94-103. Applications: Western Blotting
• Burchfield, J.G. et al. (2004) J Biol Chem 279, 18623-32. Applications: Western Blotting
• Peng, X.D. et al. (2003) Genes Dev 17, 1352-65. Applications: Western Blotting
• Ly, C. et al. (2003) Cancer Res 63, 5716-22. Applications: 2D Analysis IP Western Blotting
• Kane, S. et al. (2002) J Biol Chem 277, 22115-8. Applications: IP Western Blotting
• Lynch, D.K. and Daly, R.J. (2002) EMBO J 21, 72-82. Applications: Western Blotting
• Hermanson, O. et al. (2002) Nature 419, 934-9. Applications: Western Blotting
• Berwick, D.C. et al. (2002) J Biol Chem 277, 33895-900. Applications: Western Blotting
• Liu, M.Y. et al. (2002) Cancer Res 62, 6475-80. Applications: Western Blotting
• Wettey, F.R. et al. (2002) Science 297, 1521-5. Applications: Western Blotting
• Manning, B.D. et al. (2002) Mol Cell 10, 151-62. Applications: Western Blotting
• Fujita, N. et al. (2002) J Biol Chem 277, 28706-13. Applications: Western Blotting

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 9614 Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb
• 9621 Phospho-(Ser/Thr) PKA Substrate Antibody
• 9624 Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb
• 4060 Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb
• 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
• 4691 Akt (pan) (C67E7) Rabbit mAb
• 8112 SignalStain^® Antibody Diluent
• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.'s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.