SignalSilence? HER2/ErbB2 siRNA II

• 产品编号：CST-6283S      品牌：CST       原厂货号：6283S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

CST公司的SignalSilence® HER2/ErbB2 siRNA II能够通过RNA干扰的方法特异阻断HER2/ErbB2的表达，本方法通过向细胞内输入双链RNA选择性地对基因表达进行沉默。所有的SignalSilence® siRNA产品均经过严格的内部测试并在免疫印迹检测中显示能够降低靶蛋白的表达。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建议使用100 nM HER2/ErbB2 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。原癌基因ErbB2 (HER2)编码185kDa的受体样跨膜糖蛋白，该蛋白具有固有的酪氨酸激酶的活性 (1)。ErbB2的配体目前尚不清楚，但在高表达的ErbB2与其它ErbB家族成员形成异源二聚体时，即使缺少配体，其酶活性仍能被激活(2)。将近40%的人类乳腺癌中能够检测到ErbB2基因的扩增及其产物的过量表达。泛素连接酶c-Cbl结合在ErbB2的Tyr1112上能够导致ErbB2聚泛素化并促进其降解(4)。ErbB2是乳腺癌等多种癌肿的关键治疗靶点，通过c-Cbl介导的蛋白水解途径靶向调控ErbB2的降解是一种潜在的治疗手段。磷酸化ErbB2的激酶位点Tyr877（同源于pp60c-Src 的Tyr416）能调控ErbB2的生物活性。ErbB2的自磷酸化位点是Tyr1248和 Tyr1221/1222，这些位点的磷酸化将ErbB2与Ras-Raf-MAP激酶的信号转导途径联系起来(1,5)。

1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.

原厂资料：

Description

SignalSilence® HER2/ErbB2 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit HER2/ErbB2 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 HER2/ErbB2 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

The ErbB2 (HER2) proto-oncogene encodes a 185 kDa transmembrane, receptor-like glycoprotein with intrinsic tyrosine kinase activity (1). While ErbB2 lacks an identified ligand, ErbB2 kinase activity can be activated in the absence of a ligand when overexpressed and through heteromeric associations with other ErbB family members (2). Amplification of the ErbB2 gene and overexpression of its product are detected in almost 40% of human breast cancers (3). Binding of the c-Cbl ubiquitin ligase to ErbB2 at Tyr1112 leads to ErbB2 poly-ubiquitination and enhances degradation of this kinase (4). ErbB2 is a key therapeutic target in the treatment of breast cancer and other carcinomas and targeting the regulation of ErbB2 degradation by the c-Cbl-regulated proteolytic pathway is one potential therapeutic strategy. Phosphorylation of the kinase domain residue Tyr877 of ErbB2 (homologous to Tyr416 of pp60c-Src) may be involved in regulating ErbB2 biological activity. The major autophosphorylation sites in ErbB2 are Tyr1248 and Tyr1221/1222; phosphorylation of these sites couples ErbB2 to the Ras-Raf-MAP kinase signal transduction pathway (1,5).

1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.

注意事项：

Storage: HER2/ErbB2 siRNA II is supplied in RNAse-free water.
Aliquot and store at -20ºC.