Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (HRP Conjugate) #6950

Applications Reactivity Sensitivity Isotype
W All Endogenous Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (HRP 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.

Source / Purification

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

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and HeLa cells, untreated or treated with Calyculin A #9902 (0.1 µM, 45 min), using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (HRP Conjugate).

Description

This Cell Signaling Technology antibody is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The HRP conjugated antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb #9614.

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

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