Product Pathways - Motif Antibodies

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

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

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected

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 rabbits with phospho-Akt substrate peptides (KLH-coupled). 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 including GSK-3, Bad and caspase-9 are known, many important substrates await discovery. Akt phosphorylates substrates only at serine/threonine in a conserved motif characterized by arginine 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. et al. (1997) Annu. Rev. Biochem. 66, 807-822.
2. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol. 200, 62-82.
3. Marte, B.M. and Downward, J. (1997) Trends Biochem. Sci. 22, 355-358.
4. Jiang, B. H. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 1749-1753.
5. Scheid, M.P. and Woodgett, J.R. (2000) Curr. Biol. 10, R191-R194.
6. Alessi, D. R. et al. (1996) FEBS Lett. 399, 333-338.

Application References

• Lynch, D.K. and Daly, R.J. (2002) PKB-mediated negative feedback tightly regulates mitogenic signalling via Gab2. EMBO J. 21, 72-82. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Hermanson, O. et al. (2002) N-CoR controls differentiation of neural stem cells into astrocytes. Nature 419, 934-939. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Peng, X. et al. (2003) Dwarfism, impaired skin development, skeletal muscle atrophy, delayed bone development, and impeded adipogenesis in mice lacking Akt1 and Akt2. Genes Dev. 17, 1352-1365. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Burchfield, J. G. et al. (2004) Akt mediates insulin-stimulated phosphorylation of Ndrg2: evidence for cross-talk with protein kinase C theta. J. Biol. Chem. 279, 18623-18632. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Wada, S. et al. (2005) Phosphorylation of candida glabrata ATP-binding cassette transporter Cdr1p regulates drug efflux activity and ATPase stability. J. Biol. Chem. 280, 94-103. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Ly, C. et al. (2003) Bcr-Abl kinase modulates the translation regulators ribosomal protein S6 and 4E-BP1 in chronic myelogenous leukemia cells via the mammalian target of rapamycin. Cancer Res. 63, 5716-5722. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: 2D Analysis IP Western Blotting
• Berwick, D. C. et al. (2002) The identification of ATP-citrate lyase as a protein kinase B (Akt) substrate in primary adipocytes. J. Biol. Chem. 277, 33895-33900. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Liu, M. Y. et al. (2002) 14-3-3 interacts with the tumor suppressor tuberin at Akt phosphorylation site(s). Cancer Res. 62, 6475-6480. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Wettey, F. R. et al. (2002) Controlled elimination of clathrin heavy-chain expression in DT40 lymphocytes. Science 297, 1521-1525. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Kane, S. et al. (2002) A method to identify serine kinase substrates. Akt phosphorylates a novel adipocyte protein with a Rab GTPase-activating protein (GAP) domain. J. Biol. Chem. 277, 22115-22118. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: IP Western Blotting
• Manning, B. D. et al. (2002) Identification of the tuberous sclerosis complex-2 tumor suppressor gene product tuberin as a target of the phosphoinositide 3-kinase/Akt pathway. Mol. Cell 10, 151-162. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting
• Fujita, N. et al. (2002) Akt-dependent phosphorylation of p27Kip1 promotes binding to 14-3-3 and cytoplasmic localization. J. Biol. Chem. 277, 28706-28713. This article references the use of Phospho-(Ser/Thr) Akt Substrate Antibody in the following applications: Western Blotting

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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 commerical licensing terms please contact CST Business Development at cbunker@cellsignal.com.