E2213
Clone Type
PolyclonalHost
RabbitSource
HumanIsotype
IgGGrade
Affinity PurifiedApplications
E IC IF WBCrossreactivity
Dr Hu Mk Mo RtGene ID
1938Shipping Temp
Blue IceStorage Temp
-20°CNotes
Preservative Free
Rabbit Anti-eEF2 (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.
Applications
Suitable for use in ELISA, Western Blot, Immunocytochemistry and Immunofluorescence. Other applications not tested.
Recommended Dilution
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. Immunoflourescence (IF-IC): 1:100 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 at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic peptide corresponding to residues mapping at the N-terminus 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 total human eEF2 independent of phosphorylation at ~95kD. Species Crossreactivity: mouse, rat, monkey, D. melanogaster. Species sequence homology: 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. and Proud, C.G. (1993) Biochem. J. 293:31-34. 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., 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, 719-739.USBio References
No references available