Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (Alexa Fluor? 647 Conjugate)

• 产品编号：CST-5123S      品牌：CST       原厂货号：5123S
• 产品分类：抗体 > 一抗 > 染料标记抗体
• 应用分类：

描述：

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb 兔单抗(Alexa Fluor® 647 Conjugate)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。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。这个抗体以F/P 比值为2-5在最优条件偶联Alexa Fluor® 647。Alexa Fluor® 647染料可最大程度被红光激发(e.g. 633 nm He-Ne激光器)。偶联Alexa Fluor® 647染料的抗体能产生峰值为665 nm的远红外荧光发射。该CST抗体偶联Alexa Fluor® 647荧光染料，并用直标法流式细胞术在内部检测人类细胞。未接合的抗体Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗 #2855可与人类、小鼠、大鼠和猴子的磷酸化4E-BP1 (Thr37/46)蛋白发生反应。CST公司认为磷酸化4E-BP1 (Thr37/46) (236B4) Rabbit mAb兔单抗 (Alexa Fluor® 488接合)也将能识别这些物种磷酸化4E-BP1蛋白。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.

原厂资料：

Specificity / Sensitivity

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (Alexa Fluor® 647 Conjugate) 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.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr37 and Thr46 of mouse 4E-BP1. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-5.The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission with a peak at 665 nm.

Description

This Cell Signaling Technology (CST) antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody, Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855, reacts with Phospho-4E-BP1 (Thr37/46) from human, mouse, rat and monkey. CST expects that phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (Alexa Fluor® 647 Conjugate) will also recognize Phospho-4E-BP1 in these species.

Background

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).

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.

注意事项：

Storage: Supplied in PBS (pH 7.2), less than 0.1% sodium
azide, 2 mg/ml BSA. Store at 4°C. Protect from light. Do not
freeze.