SignalSilence? ERCC1 siRNA I

• 产品编号：CST-8896S      品牌：CST       原厂货号：8896S
• 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® ERCC1 siRNA 帮助研究人员使用RNA干扰技术特异性抑制ERCC1表达，该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试，western分析显示能够能够降低目标蛋白的表达。寡核苷酸合成经过了三苯甲基的逐碱基分析以确保耦合效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。CST建议转染100 nM SignalSilence® ERCC1 siRNA I，48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题，可以随时咨询CST。 每管试剂能进行100次转染，该转染次数以总体积为300μl 的1个24孔板孔中siRNA终浓度100 nM来进行计算。所有活细胞都有DNA损伤修复功能以调控DNA损伤。核苷酸切除修复（NER）出现在大片段的螺旋-扭曲损伤时，例如由UV或某种化学物质引发的损伤（1）。Excision Repair Cross Complementing 1 (ERCC1)与XPF构成复合物，作为5’内切酶切除掉发生损伤的部位（2）。ERCC1-XPF还在DNA链间交联（ICLs）（3）和双链损伤时（4）发挥足以。研究证明ERCC1的表达与存活率相关，也在一些人类癌症疾病，包括肺小细胞癌（NSCLC）对化疗的的反应过程相关（5,6）。

1. Shuck, S.C. et al. (2008) Cell Res 18, 64-72.
2. McDaniel, L.D. and Schultz, R.A. (2008) Adv Exp Med Biol 637, 65-82.
3. Niedernhofer, L.J. et al. (2004) Mol Cell Biol 24, 5776-87.
4. Ahmad, A. et al. (2008) Mol Cell Biol 28, 5082-92.
5. Zheng, Z. et al. (2007) N Engl J Med 356, 800-8.
6. Gossage, L. and Madhusudan, S. (2007) Cancer Treat Rev 33, 565-77.

原厂资料：

Description

SignalSilence® ERCC1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit ERCC1 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 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® ERCC1 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. 
 Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

DNA repair systems operate in all living cells to manage a variety of DNA lesions. Nucleotide excision repair (NER) is implemented in cases where bulky helix-distorting lesions occur, such as those brought about by UV and certain chemicals (1). Excision Repair Cross Complementing 1 (ERCC1) forms a complex with XPF, which acts as the 5’ endonuclease required to excise the lesion (2). ERCC1-XPF is also required for repair of DNA interstrand crosslinks (ICLs) (3) and involved in repair of double strand breaks (4). Research studies have shown that expression of ERCC1 is related to survival rate and response to chemotherapeutic drugs in several human cancers including non-small cell lung cancer (NSCLC) (5,6).

1. Shuck, S.C. et al. (2008) Cell Res 18, 64-72.
2. McDaniel, L.D. and Schultz, R.A. (2008) Adv Exp Med Biol 637, 65-82.
3. Niedernhofer, L.J. et al. (2004) Mol Cell Biol 24, 5776-87.
4. Ahmad, A. et al. (2008) Mol Cell Biol 28, 5082-92.
5. Zheng, Z. et al. (2007) N Engl J Med 356, 800-8.
6. Gossage, L. and Madhusudan, S. (2007) Cancer Treat Rev 33, 565-77.

注意事项：

Storage: ERCC1 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20°C