Technical Data

E2213-01
Clone Type
Polyclonal
Host
Rabbit
Source
Human
Isotype
IgG
Grade
Affinity Purified
Applications
WB
Crossreactivity
Ch Hm Hu Mk Mo Rt
Gene ID
1938
Shipping Temp
Blue Ice
Storage Temp
-20°C
Notes
Preservative Free
Rabbit Anti-eEF2, phosphorylated (Thr56) (Elongation Factor 2, Eukaryotic EF2)

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).

Applications
Suitable for use in Western Blot. Other applications not tested.
Recommended Dilutions
Western Blot: 1:1000; Incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding Thr56 of human eEF2.
Form
Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.
Purity
Purified by Protein A and peptide affinity chromatography.
Specificity
Recognizes endogenous levels of human eEF2 at ~95kD only when phosphorylated at Thr56. Does not recognize eEF2 phosphorylated at other sites. Species Crossreactivity: mouse, rat, monkey, chicken and hamster.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Nairn, A.C. and Palfrey, H.C. (1987) J. Biol. Chem. 262:17299–17303. 2. Ryazanov, A.G. et al. (1988) Nature 334:170–173. 3. Carlberg, U. et al. (1990) Eur. J. Biochem. 191:639–645. 4. Redpath, N.T. et al. (1993) Eur. J. Biochem. 213:689–699. 5. Nairn, A.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82:7939–7943. 6. Palfrey, H.C. et al. (1987) J. Biol. Chem. 262: 9785–9792. 7. Redpath, N.T. and Proud, C.G. (1993) Biochem. J. 293:31–34. 8. Diggle, T. et al. (1998) Biochem. J. 336:525–529. 9. Hovland, R. et al. (1999) FEBS Lett. 444:97–101. 10. Proud, C. (2000) Translational Control of Gene Expression. Cold Spring Harbor Laboratory Press, NY, 719–739
USBio References
No references available
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