Cell Signaling Technology

Product Pathways - Motif Antibodies

Immobilized Phospho-(Ser/Thr) Akt Substrate Antibody #9619

Applications Reactivity Source
IP All Rabbit

Applications Key:  IP=Immunoprecipitation
Reactivity Key: All=All species expected

Specificity / Sensitivity

Immobilized Phospho-(Ser/Thr) Akt Substrate Antibody is useful for immunoprecipitation of phosphorylated Akt substrate proteins. This antibody recognizes proteins containing phospho-Ser/Thr preceded by Arg or Lys at positions -5 and -3. It shows some cross-reactivity with phospho-Ser/Thr preceded by Arg or Lys at positions -3 and -2. It does not cross-react with other phospho-Ser/Thr-containing motifs. The antibody is immobilized by conjugation of carbohydrates to cross-linked agarose hydrazide beads. (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 a phospho-Akt substrate peptide (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or PDGF-treated (50 ng/ml for 15 minutes) compared with proteins immunoprecipitated or supernatants collected after the immunoprecipitations from the same lysates, using Immobilized Phospho-(Ser/Thr) Akt Substrate Antibody. Western blot was performed using Phospho-(Ser/Thr) Akt Substrate Antibody #9611.

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

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Companion Products

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.

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