Cell Signaling Technology

Product Pathways - Translational Control

eEF2k Antibody #3692

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC H R Mk Endogenous 105 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

eEF2k Antibody detects endogenous levels of total eEF2k independent of phosphorylation.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the N-terminus of human eEF2k. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from C6, 293, HeLa and SW-13 cells using eEF2k Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using eEF2k Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Eukaryotic elongation factor 2 kinase (eEF2k) phosphorylates and inactivates eEF2, resulting in the inhibition of peptide-chain elongation (1). eEF2k is normally dependent on Ca2+ ions and calmodulin (2,3). It can be activated by PKA in response to elevated cAMP levels (4-6), which are generally increased in stress- or starvation-related conditions. eEF2k can also be regulated in response to a wide range of stimuli that promote cell growth and protein synthesis. This involves the phosphorylation of eEF2k by p90RSK and p70 S6 kinase at Ser366 or by SAPK4/p38delta at Ser359, leading to the inactivation of eEF2k (7,8), which facilitates the dephosphorylation of eEF2, and thus promotes translation.

  1. Ryazanov, A.G. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 4884-4889.
  2. Nairn, A.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82, 7839-7943.
  3. Palfrey, H.C. et al. (1987) J. Biol. Chem. 262, 9785-9792.
  4. Redpath, N.T. et al. (1993) Biochem. J. 293, 31-34.
  5. Diggle, T.A. et al. (1998) Biochem. J. 336, 525-529.
  6. Hovland, R. et al. (1999) FEBS Lett. 444, 97-101.
  7. Wang, X. et al. (2001) EMBO J. 20, 4370-4379.
  8. Knebel, A. et al. (2001) EMBO J. 20, 4360-4369.

Application References

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Companion Products

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.

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