Cell Signaling Technology

Product Pathways - Protein Translation

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855

4E-BP   4E-BP1   4EBP   4EBP1   PHAS   PHAS-1  

No. Size Price
2855L 300 µl ( 30 western blots ) ¥8,992.00 现货查询 购买询价 防伪查询
2855S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价 防伪查询
2855T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价 防伪查询
2855 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey,D. melanogaster, Endogenous 15 to 20 Rabbit IgG
IHC-P 1:1600
F 1:50
IF-IC 1:200

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb detects endogenous levels of 4E-BP1 only when phosphorylated at Thr37 and/or Thr46. This antibody may cross-react with 4E-BP2 and 4E-BP3 when phosphorylated at equivalent sites.

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗可检测仅在Thr37或Thr46位点磷酸化的内源性4E-BP1蛋白水平。该抗体可以与在等效位点磷酸化的4E-BP2和4E-BP3蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr37 and Thr46 of mouse 4E-BP1.

通过人工合成小鼠4E-BP1 蛋白Thr37和Thr46位点周围序列相应的多肽片段去免疫动物从而制备单克隆抗体。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.

使用Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗,免疫组化分析人类淋巴瘤组织石蜡切片。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells untreated (left) or LY294002-treated (right)).

使用Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗,免疫组化分析SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (未处理(左图) 和LY294002处理 (右图)的石蜡包埋的LNCaP细胞。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue), using Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb compared to a nonspecific negative control antibody (red).

与阴性非特异的对照抗体(红色)比较,使用Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb兔单抗,流式细胞术分析Jurkat细胞,细胞分为未处理组(绿色)或LY294002、Wortmannin和U0126处理组(蓝色)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.

使用Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗,免疫组化分析人类结肠癌组织石蜡切片。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb in the presence of control peptide (left) or Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052 (right).

使用Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗,免疫组化分析人类结肠癌组织石蜡切片,其分别孵育对照多肽(左图)和Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052 (右图)。



Confocal immunofluorescent analysis of HeLa cells treated with LY294002 (left) or 20% serum (right) and labeled with Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

使用Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗(绿色),共聚焦免疫荧光观察Miwi蛋白在HeLa细胞中定位,细胞分别使用LY294002处理(左图)或20%血清处理(右图)。Alexa Fluor® 555 phalloidin标记微丝蛋白(红色)。

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells using 4E-BP1 Antibody #9452 (upper) and Phospho-4E-BP1 (Thr37/46) Antibody #2855 (lower). The cells were starved for 24 hours in serum-free medium and underwent a 1 hour amino acid deprivation. Amino acids were replenished for 1 hour. Cells were then either untreated (-) or treated with 100 nM insulin (+) for 30 minutes.

使用4E-BP1 Antibody #9452 (上图)和Phospho-4E-BP1 (Thr37/46) Antibody #2855 (下图),免疫印迹(Western Blot)分析293 T细胞系裂解物。细胞在无血清饥饿培养24小时,接着氨基酸缺乏培养1小时,然后再加入氨基酸培养1小时。最后,细胞加入100 nM胰岛素 (+)或不加(-)处理30分钟。



Confocal immunofluorescent analysis of 293 cells, expressing either non-targeting shRNA (top) or shRNA targeting 4E-BP1/2 (bottom), using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). To confirm phospho-specificity, cells were treated with an inhibitor cocktail consisting of LY294002 #9901, U0126 #9903, and Rapamycin #9904 (50 μM; 10 μm; 10 nM; 2 hr) (left), stimulated with insulin (100 nM, 30 min; middle), or processed with λ-phosphatase following insulin stimulation (right). Red = Propidium Iodide (PI)/RNase Staining Solution (#4087).


Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR at Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).

翻译抑制蛋白4E-BP1 (又称 PHAS-1)通过结合翻译起始因子eIF4E蛋白从而抑制帽子结构依赖的翻译。4E-BP1的过磷酸化干扰上述结合并且导致激活帽子结构依赖的翻译(1)。PI3 kinase/Akt通路和FRAP/mTOR激酶通路都调节4E-BP1蛋白的激活(2,3)。在体内4E-BP1多个位点残基被磷酸化(4)。通过FRAP/mTOR蛋白使4E-BP1在Thr37和Thr46位点磷酸化不能阻止它eIF4E结合,但这被认为是为随后在4E-BP1的Ser65和Thr70位点磷酸化进行准备(5)。

  1. Pause, A. et al. (1994) Nature 371, 762-7.
  2. Brunn, G.J. et al. (1997) Science 277, 99-101.
  3. Gingras, A.C. et al. (1998) Genes Dev 12, 502-13.
  4. Fadden, P. et al. (1997) J Biol Chem 272, 10240-7.
  5. Gingras, A.C. et al. (1999) Genes Dev 13, 1422-37.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

U.S. Patent No. 5,675,063.

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