WB
H M R Hm Mk C
Endogenous
95
Rabbit
#P13639
1938
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Hamster, Monkey, Chicken
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr56 of human eEF2. Antibodies are purified by protein A and peptide affinity chromatography.
Background
Eukaryotic elongation factor 2 (eEF2) catalyzes the translocation of peptidyl-tRNA from the A site to the P site on the ribosome. It has been shown that phosphorylation of eEF2 at threonine 56 by eEF2 kinase inhibits its activity (1-4). eEF2 kinase is normally dependent on Ca2+ ions and calmodulin (5,6). eEF2 kinase can also be activated by PKA in response to elevated cAMP levels (7-9), which are generally increased in stress- or starvation-related conditions. A variety of treatments known to raise intracellular Ca2+ or cAMP levels have been shown to result in increased phosphorylation of eEF2, and thus to inhibit peptide-chain elongation. The inactive phosphorylated eEF2 can be converted to its active nonphosphorylated form by a protein phosphatase, most likely a form of protein phosphatase-2A (PP-2A). Insulin, which activates protein synthesis in a wide range of cell types, induces rapid dephosphorylation of eEF2 through mTOR signaling and may involve modulation of the activity of the PP-2A or the eEF2 kinase or both (10).
- Nairn, A.C. and Palfrey, H.C. (1987) J. Biol. Chem. 262, 17299-17303.
- Ryazanov, A.G. et al. (1988) Nature 334, 170-173.
- Carlberg, U. et al. (1990) Eur. J. Biochem. 191, 639-645.
- Redpath, N.T. et al. (1993) Eur. J. Biochem. 213, 689-699.
- Nairn, A.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82, 7939-7943.
- Palfrey, H.C. et al. (1987) J. Biol. Chem. 262, 9785-9792.
- Redpath, N.T. and Proud, C.G. (1993) Biochem. J. 293, 31-34.
- Diggle, T. et al. (1998) Biochem. J. 336, 525-529.
- Hovland, R. et al. (1999) FEBS Lett. 444, 97-101.
- Proud, C. (2000) Translational Control of Gene Expression. Cold Spring Harbor Laboratory Press, NY, 719-739.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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