SignalSilence? Met siRNA I

• 产品编号：CST-6618S      品牌：CST       原厂货号：6618S
• 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® Met siRNA I能够通过RNA干扰的方法特异阻断Met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 Met siRNA I48 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。Met作为肝细胞生长因子 (HGF, 亦作扩散因子) 的高亲和性酪氨酸激酶受体是一个二硫键连接的异二聚体，含有一个45 kDa的 α-亚基和一个145 kDa的 β-亚基 (1,2)。α-亚基和β-亚基的氨基端形成胞外区，β-亚基的其余部分跨细胞膜并且含有一个具有激酶活性的胞内区。Met和HGF相互作用会导致多个酪氨酸位点的自磷酸化，继而招募下游信号分子Gab1, c-Cbl和 PI3 K激酶（3）。这一系列基础性事件对于Met激酶相关的所有生物学功能十分重要。胞内Tyr1003位点的磷酸化对于Met的泛素化和降解是必须的（4）。激酶结构域Tyr1234/1235的磷酸化对于Met的激酶活性十分关键。胞内 Tyr1349的磷酸化为Met提供了Gab1 的直接结合位点(5)。在肾、结肠和乳腺癌中发现有Met水平和/或激酶活性的改变，因此Met是肿瘤治疗和诊断的一个靶点（6,7）。

1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

原厂资料：

Description

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

Met, a high affinity tyrosine kinase receptor for hepatocyte growth factor (HGF, also known as scatter factor) is a disulfide-linked heterodimer made of 45 kDa α- and 145 kDa β-subunits (1,2). The α-subunit and the amino-terminal region of the β-subunit form the extracellular domain. The remainder of the β-chain spans the plasma membrane and contains a cytoplasmic region with tyrosine kinase activity. Interaction of Met with HGF results in autophosphorylation at multiple tyrosines, which recruit several downstream signaling components, including Gab1, c-Cbl, and PI3 kinase (3). These fundamental events are important for all of the biological functions involving Met kinase activity. The addition of a phosphate at cytoplasmic Tyr1003 is essential for Met protein ubiquitination and degradation (4). Phosphorylation at Tyr1234/1235 in the Met kinase domain is critical for kinase activation. Phosphorylation at Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon, and breast. Thus, Met is an attractive cancer therapeutic and diagnostic target (6,7).

1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

注意事项：

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