Cell Signaling Technology

Product Pathways - Protein Translation

Phospho-eIF4B (Ser406) Antibody #5399

4B   EIF   eIF-4B   eif4   EIF4B   IF4B   translation initiation factor  

No. Size Price
5399S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
5399 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 80 Rabbit
IP 1:50
IF-IC 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Rat,

Specificity / Sensitivity

Phospho-eIF4B (Ser406) Antibody detects endogenous levels of eIF4B protein only when phosphorylated at Ser406.

Phospho-eIF4B (Ser406) Antibody检测仅在Ser406位点磷酸化的内源性eIF4B蛋白水平。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser406 of human eIF4B protein. Antibodies are purified by protein A and peptide affinity chromatography.

通过人工合成人源eIF4B蛋白Ser406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 NIH/3T3 cells, untreated or λ phosphatase treated, using Phospho-eIF4B (Ser406) Antibody (upper) or eIF4B Antibody #3592 (lower).

使用Phospho-eIF4B (Ser406) Antibody (上图)和eIF4B Antibody #3592 (下图),免疫印迹(Western Blot)分析在NIH/3T3细胞系中磷酸化eIF4B (Ser406)和eIF4B蛋白水平,细胞分为对照组和λ磷酸酶处理组。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (upper) or λ phosphatase treated (lower), using Phospho-eIF4B (Ser406) Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).

使用Phospho-eIF4B (Ser406) Antibody(绿色)标记,共聚焦免疫荧光观察磷酸化eIF4B (Ser406)蛋白在HeLa细胞中定位,细胞分为对照组和λ磷酸酶处理组。DY-554 phalloidin (红色)标记微丝蛋白。蓝色伪彩= AQ5® #4084 (DNA荧光染料)。

Background

Eukaryotic initiation factor 4B (eIF4B) is thought to assist the eIF4F complex in translation initiation. In plants, eIF4B is known to interact with the poly-(A) binding protein, increasing its poly-(A) binding activity (1). Heat shock and serum starvation cause dephosphorylation of eIF4B at multiple sites with kinetics similar to those of the corresponding inhibition of translation, while phosphorylation of eIF4B following insulin treatment correlates well with an observed increase in translation (2-5). Multiple kinases, including p70 S6 kinase, can phosphorylate eIF4B in vitro, and at least one serum-inducible eIF4B phosphorylation site is sensitive to rapamycin and LY294002 (6). Recently Ser406 was identified as a novel phosphorylation site regulated by mitogens (7), and the phosphorylation of this site is dependent on the MEK and mTOR activity (7). This phosphorylation is shown to be essential for the translational activity of eIF4B (7).

真核生物起始因子4B (eIF4B)被认为在蛋白质翻译起始阶段协助eIF4F复合物。在植物中,已知eIF4B蛋白和poly-(A)结合蛋白相互作用,从而增加它与poly-(A)结合活性(1)。虽然eIF4B在胰岛素刺激下发生磷酸化和蛋白质翻译增加保持很大的相关性,但是热休克和血清饥饿能够引起eIF4B在多个位点去磷酸化,这类似一些翻译抑制的动力学特征(2-5)。许多激酶包括p70 S6 激酶能够在体外使eIF4B磷酸化,同时至少一个血清诱导的eIF4B磷酸化位点对rapamycin和LY294002敏感(6)。最近研究证明Ser406位点是通过有丝分裂素调节的新型磷酸化位点(7),同时这个磷酸化位点是依赖于MEK和mTOR通路的活化(7)。因此,这个位点的磷酸化对eIF4B翻译活性起着至关重要的作用(7)。

  1. Le, H. et al. (1997) J. Biol. Chem. 272, 16247-16255.
  2. Duncan, R.F. and Hershey, J.W. (1989) J. Cell Biol. 109, 1467-1481.
  3. Duncan, R.F. and Hershey, J.W. (1984) J. Biol. Chem. 259, 11882-11889.
  4. Duncan, R. and Hershey, J.W. (1985) J. Biol. Chem. 260, 5493-5497.
  5. Manzella, J.M. et al. (1991) J. Biol. Chem. 266, 2383-2389.
  6. Gingras, A.C. et al. (2001) Genes Dev. 15, 807-826.
  7. van Gorp, A.G. et al. (2009) Oncogene 28, 95-106.

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