antibody

eEF2, phosphorylated (Thr56) (Elongation Factor 2, Eukaryotic EF2)

E2213-01

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

eEF2, phosphorylated (Thr56) (Elongation Factor 2, Eukaryotic EF2)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

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 ELISA and 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.

Ch Hm Hu Mk Mo Rt

Synthetic phosphopeptide corresponding to residues surrounding Thr56 of human eEF2.

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.

Purified by Protein A and peptide affinity chromatography.

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.

Not Determined

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: Not Determined 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. Immunogen: Synthetic phosphopeptide corresponding to residues surrounding Thr56 of human eEF2. 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. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

E WB

-20°C

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

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