Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (Sepharose Bead Conjugate) #9646

Applications Reactivity Sensitivity Isotype
IP All Endogenous Rabbit IgG

Applications Key:  IP=Immunoprecipitation
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Phospho-(Ser/Thr) Akt Substrate Motif (RXXS*/T*) (110B7) Rabbit mAb (Sepharose Bead Conjugate) recognizes peptides and proteins containing phospho-serine/threonine preceded by arginine at the -3 position. There is some preference observed for peptides that contain phospho-serine/threonine preceded by arginine at both positions -5 and -3. (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

Monoclonal antibody is produced by immunizing animals with synthetic phospho-Akt substrate peptides.

IP

IP

Immunoprecipitation of NIH/3T3 cell lysates, untreated or PDGF-treated, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (Sepharose Bead Conjugate) compared to supernatants collected after immunoprecipitation from the same lysate. The western blot was probed using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb #9614.

Description

This Cell Signaling Technology antibody is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated sepharose beads. Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (Sepharose Bead Conjugate) is useful for the immunoprecipitation of phosphorylated Akt substrate proteins.

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

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Companion Products

Produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and U.S.S.N. 11/476,277) from Epitomics, Inc.

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

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