SignalSilence? 4E-BP1 siRNA I

• 产品编号：CST-6414S      品牌：CST       原厂货号：6414S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司的SignalSilence® 4E-BP1 siRNA允许研究者通过RNA干扰去特异性抑制4E-BP1表达，这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence^® siRNA产品都是经过内部严格检测的，并且通过Western blot 分析证明确实能够减少目的蛋白的表达。通过三苯甲基分析每个碱基以监测寡核苷酸的合成，确保合适的配对效率。随后寡核苷酸通过亲和固相萃取法纯化。退火的RNA双链通过质谱分析来证实其精确的组成。每一批产品都通过质谱分析与前面的产品进行比较，来保证不同批次之间的最大一致性。CST公司推荐在细胞裂解前48到72小时转染100 nM SignalSilence® 4E-BP1 siRNA I 。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题，请随时联系CST。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：

Description

SignalSilence® 4E-BP1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit 4E-BP1 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® 4E-BP1 siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

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: SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.