SignalSilence? ATR siRNA II

• 产品编号：CST-6289S      品牌：CST       原厂货号：6289S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

 CST的SignalSilence® ATR siRNA II允许研究人员采用RNA干扰（一种通过传递外源性的双链RNA分子到细胞中从而能选择性的沉默基因表达的方法）技术特异性的抑制ATR蛋白表达。CST所有的SignalSilence® siRNA产品都经过了严格的内部测试并采用Western分析证明其可以有效降低靶蛋白的表达。为了保证适当的结合效率，我们采用三苯甲基分析的方法逐个碱基的监测寡核苷酸的合成。随后通过亲和固相提取纯化寡核苷酸。退火的双链RNA进一步通过质谱分析来验证双链复合体的确切成分。每一批产品和前一批都要通过质谱分析进行比较以确定批次间最大限度的一致性。 CST建议使用100 nM ATR siRNA II转染48-72小时后裂解细胞。转染的具体步骤请参考转染试剂制造商提供的操作标准。使用过程中有任何问题请随时联系CST。

1. Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86.
2. Abraham, R.T. (2004) DNA Repair (Amst) 3, 883-7.
3. Shechter, D. et al. (2004) DNA Repair (Amst) 3, 901-8.
4. Vauzour, D. et al. (2007) Arch Biochem Biophys 468, 159-66.
5. Smith, J. et al. (2010) Adv Cancer Res 108, 73-112.
6. Nam, E.A. et al. (2011) J Biol Chem 286, 28707-14.
7. Liu, S. et al. (2011) Mol Cell 43, 192-202.

原厂资料：

Description

SignalSilence® ATR siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit ATR 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 ATR siRNA II 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

Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are PI3 Kinase-related kinase (PIKK) family members that phosphorylate multiple substrates on serine or threonine residues that are followed by a glutamine in response to DNA damage or replication blocks (1-3). Despite the essential role of ATR in cell cycle signaling and DNA repair processes, little is known about its activation. While there have been no published reports of phosphorylation sites on ATR, Cell Signaling Technology has produced an antibody directed against phospho-ATR (Ser428) that demonstrates in vivo and UV-induced phosphorylation of this protein. This reagent could prove to be a valuable tool for monitoring ATR activation. Proline-directed phosphorylation sites like this one are often targeted by CDKs and MAPKs and can often dramatically affect protein conformation (4,5). 

1. Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86.
2. Abraham, R.T. (2004) DNA Repair (Amst) 3, 883-7.
3. Shechter, D. et al. (2004) DNA Repair (Amst) 3, 901-8.
4. Vauzour, D. et al. (2007) Arch Biochem Biophys 468, 159-66.
5. Smith, J. et al. (2010) Adv Cancer Res 108, 73-112.
6. Nam, E.A. et al. (2011) J Biol Chem 286, 28707-14.
7. Liu, S. et al. (2011) Mol Cell 43, 192-202.

注意事项：

Storage: ATR siRNA II is supplied in RNAse-free water. Aliquot
and store at -20ºC.