Cell Signaling Technology

Product Pathways - Protein Translation

Phospho-eEF2k (Ser366) Antibody #3691

eEF2 kinase   eef2k   Eukaryotic elongation factor 2 kinase  

No. Size Price
3691S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
3691T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
3691 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Rat,Monkey, Endogenous 105 Rabbit
IP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Specificity / Sensitivity

Phospho-eEF2k (Ser366) Antibody detects endogenous levels of eEF2k only when phosphorylated at Ser366.

Phospho-eEF2k (Ser366) Antibody检测仅在Ser366位点磷酸化的内源性eEF2k蛋白水平。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser366 of human eEF2k. Antibodies are purified by protein A and peptide affinity chromatography.

通过人工合成人源eEF2k蛋白Ser366位点周围序列相应的磷酸化片段去免疫动物从而制备多克隆抗体。通过蛋白A和多肽亲和层析纯化抗体。

Western Blotting

Western Blotting

Western blot analysis of extracts from SW-13 cells (starved for 18 hours) treated with calf intestinal alkaline phosphatase (CIP) or 20% fetal bovine serum for 30 minutes, and extracts from HeLa cells (starved for 18 hours) treated with 200 nM TPA for 30 minutes, using Phospho-eEF2k (Ser366) Antibody (upper) or eEF2k Antibody #3692 (lower).

使用Phospho-eEF2k (Ser366) Antibody (上图)和eEF2k Antibody #3692 (下图),免疫印迹(Western Blot)分析在SW-13细胞和HeLa细胞中磷酸化eEF2k和eEF2k蛋白水平, SW-13(饥饿处理18小时)细胞提取物分为牛小肠碱性磷酸酶(CIP)处理30分钟组和20%胎牛血清处理30分钟组。HeLa细胞(饥饿处理18小时)的提取物是经过200 nM TPA处理30分钟。

Background

Eukaryotic elongation factor 2 kinase (eEF2k) phosphorylates and inactivates eEF2, resulting in the inhibition of peptide-chain elongation (1). eEF2k is normally dependent on Ca2+ ions and calmodulin (2,3). It can be activated by PKA in response to elevated cAMP levels (4-6), which are generally increased in stress- or starvation-related conditions. eEF2k can also be regulated in response to a wide range of stimuli that promote cell growth and protein synthesis. This involves the phosphorylation of eEF2k by p90RSK and p70 S6 kinase at Ser366 or by SAPK4/p38delta at Ser359, leading to the inactivation of eEF2k (7,8), which facilitates the dephosphorylation of eEF2, and thus promotes translation.

Eukaryotic elongation factor 2 kinase (eEF2k)使eEF2蛋白磷酸化并失活,从而导致多肽链延伸过程受到抑制(1)。通常eEF2k是一种Ca2+离子和钙调蛋白依赖性激酶(2,3)。eEF2激酶也能被PKA所激活(7-9),而PKA通常是响应因应激增加或饥饿相关条件下cAMP水平的上升(4-6)。eEF2k也能够在促细胞生长和蛋白质合成的众多刺激下被调节。这种调节分别涉及到通过p90RSK对eEF2K的磷酸化,p70 S6 kinase在Ser366位点或SAPK4/p38delta在Ser359位点对eEF2K的磷酸化,从而导致eEF2k失活(7,8)。而这有助于eEF2的去磷酸化,同时促进蛋白质翻译。

  1. Ryazanov, A.G. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 4884-4889.
  2. Nairn, A.C. et al. (1985) Proc. Natl. Acad. Sci. USA 82, 7839-7943.
  3. Palfrey, H.C. et al. (1987) J. Biol. Chem. 262, 9785-9792.
  4. Redpath, N.T. et al. (1993) Biochem. J. 293, 31-34.
  5. Diggle, T.A. et al. (1998) Biochem. J. 336, 525-529.
  6. Hovland, R. et al. (1999) FEBS Lett. 444, 97-101.
  7. Wang, X. et al. (2001) EMBO J. 20, 4370-4379.
  8. Knebel, A. et al. (2001) EMBO J. 20, 4360-4369.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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