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Technical Data Sheet

EL7351

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eEF2K Recombinant Protein
Lysate
Price
Size

$75
100 μl

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Enlarge

Recombinant human eEF2K untreated (lanes 1 & 4) and auto-phosphorylated in the presence of Ca2+ and Calmodulin (lanes 2 & 5). The eEF2K was dephosphorylated with lambda phosphatase (lanes 3 & 6). The blots were probed with anti-eEF2K (N-terminus) (lanes 1-3) or anti-eEF2K (Thr-348) (lanes 4-6). (Images provided by Dr. Kevin Dalby in the Dept. of Pharmacy at the University of Texas.)

Application
Dilution


WB
10 μl/lane



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Eukaryotic elongation factor 2 (eEF2) catalyzes the translocation of peptidyl-tRNA from the A site to the P site on the ribosome. eEF2 kinase (eEF2K) phosphorylates and inactivates eEF2, resulting in the inhibition of peptide-chain elongation. eEF2K is normally dependent on Ca2+ ions and calmodulin, and can be activated by PKA in response to elevated cAMP levels during cell stress- or starvation-related conditions. Regulation of eEF2K occurs through phosphorylation at multiple sites. Ser-78 phosphorylation is required for calmodulin binding and eEF2K activity, while phosphorylation of Ser-500 is required for Ca2+/calmodulin-independent kinase activity. Thr-348 is an autophosphorylation site that is required for kinase activity. Inhibitory phosphorylation may also regulate eEF2K, since phosphorylation at Ser-359 by SAPK4/p38delta causes inactivation of eEF2K.

Human recombinant eEF2K is a full length protein (95 kDa) expressed in E. Coli. The protein is detected by rabbit polyclonal anti-eEF2K (C-terminus) antibody (Cat.#EP4661) and anti-eEF2K (Thr-348) (Cat.#EP4411).


Background References
Ryazanov, A.G. et al. (1997) Proc Natl Acad Sci. USA. 94:4884.
Diggle, T.A., et al. (2001) Biochem J. 353(3):621.
Smith, E.M. & Proud, C.G. (2008) EMBO J. 27(7):1005.
Tavares, C.D. et al. (2012) Biochemistry. 51(11):2232.
Buffer and Storage
eEF2K recombinant protein lysate is supplied at a concentration of 5 ng/µl in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. Stable for 1 year.
imageRelated Products
EK6910
eEF2K Phospho-Regulation Antibody Sampler Kit

EP4391
eEF2K (Ser-78), phospho-specific Rabbit Polyclonal

EP4411
eEF2K (Thr-348), phospho-specific Rabbit Polyclonal

EP4431
eEF2K (Ser-359), phospho-specific Rabbit Polyclonal

EP4451
eEF2K (Ser-500), phospho-specific Rabbit Polyclonal

EP4661
eEF2K (C-terminus) Rabbit Polyclonal

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