Our eEF2K (Thr-348) rabbit polyclonal phosphospecific primary antibody from PhosphoSolutions is produced in-house. It detects human, mouse, and rat eEF2K (Thr-348) and is antigen affinity purified. It is great for use in WB.
Recombinant human eEF2K untreated (lanes 1 & 4) and auto-phosphorylated in the presence of Ca2+ and Calmodulin (lanes 2 & 5). In some lanes, the eEF2K was dephosphorylated with lambda phosphatase (lanes 3 & 6). The blots were probed with rabbit polyclonal anti-eEF2K (N-terminus) (lanes 1-3) or anti-eEF2K (Thr-348) (lanes 4-6). (Images provided by the laboratory of Dr. Kevin Dalby in the Dept. of Pharmacy at the University of Texas at Austin.)
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/p38δ causes inactivation of eEF2K. Thus, multisite phospho-regulation of eEF2K may be important for proper control of eEF2K activity and protein translation.
Antigen Affinity Purified
Polyclonal
IgG
WB
Rabbit
EEF2K
105
Phospho-eEF2K (Thr-348) synthetic peptide (coupled to carrier protein) corresponds to amino acids surrounding Thr-348 in human eEF2K. This sequence is well conserved in rat and mouse eEF2K, and has low homology to other proteins.
Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
Liquid
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
WB: 1:1000
Unconjugated
The antibody detects a 105 kDa* protein corresponding to the molecular mass of eEF2K on SDS-PAGE immunoblots of human recombinant eEF2K autophosphorylated in the presence of Ca2+ and Calmodulin. This reactivity is removed after lambda phosphatase treatment.
Phosphorylated
Thr-348
Western blots performed on each lot.
For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.
United States
After date of receipt, stable for at least 1 year at -20°C.
Piserchio, A, et al. 2021. Structural dynamics of the complex of calmodulin with a minimal functional construct of eukaryotic elongation factor 2 kinase and the role of Thr348 autophosphorylation. Protein science : a publication of the Protein Society, 1221-1234.
Tavares, C.D., et al. 2017. Signal Integration at Elongation Factor 2 Kinase: THE ROLES OF CALCIUM, CALMODULIN, AND SER-500 PHOSPHORYLATION. Journal of Biological Chemistry, 2032-2045.
Tavares, C.D., et al. 2012. Calcium/calmodulin stimulates the autophosphorylation of elongation factor 2 kinase on Thr-348 and Ser-500 to regulate its activity and calcium dependence. Biochemistry, 2232-2245.
Fuchs, T, et al. 2017. Disinhibition of somatostatin-positive GABAergic interneurons results in an anxiolytic and antidepressant-like brain state. Molecular psychiatry, 920-930.
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