SignalSilence? Ezrin siRNA I

• 产品编号：CST-6475S      品牌：CST       原厂货号：6475S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

来自Cell Signaling Technology (CST)的SignalSilence^® RMP siRNA I可以帮助研究者通过RNA干扰特异性地抑制RMP的表达，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推荐使用100 nM SignalSilence^® Ezrin siRNA I进行转染，48到72小时后对细胞进行裂解。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。ezrin, radixin, 和moesin (ERM)蛋白功能作为细胞质膜和肌动蛋白细胞骨架的连接者，并且该蛋白涉及细胞粘附、胞膜边缘波动(membrane ruffling)和微绒毛形成 (1)。ERM蛋白参与分子内或分子间的氨基和羧基端结构域的相互作用，该蛋白是以非活性的细胞基质单体或二聚体存在 (2)。ERM蛋白在羧基端残基(ezrin蛋白Thr567位点、radixin蛋白Thr564位点、moesin蛋白Thr558 位点) 磷酸化可干扰氨基端和羧基端的关联，这可能在调节ERM蛋白的形成和功能中起着重要作用 (3,4)。ezrin蛋白Thr567位点磷酸化对于细胞骨架的重组和致癌基因诱导的转化可能起着关键角色(5)。基于生长因子刺激，Ezrin蛋白也在酪氨酸残基发生磷酸化。在上皮细胞分化期间，ezrin蛋白Tyr353位点的磷酸化传递一个生存信号(6)。

1. Tsukita, S. and Yonemura, S. (1999) J Biol Chem 274, 34507-10.
2. Mangeat, P. et al. (1999) Trends Cell Biol 9, 187-92.
3. Matsui, T. et al. (1998) J Cell Biol 140, 647-57.
4. Gautreau, A. et al. (2000) J Cell Biol 150, 193-203.
5. Tran Quang, C. et al. (2000) EMBO J 19, 4565-76.
6. Gautreau, A. et al. (1999) Proc Natl Acad Sci U S A 96, 7300-5.

原厂资料：

Description

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

The ezrin, radixin, and moesin (ERM) proteins function as linkers between the plasma membrane and the actin cytoskeleton and are involved in cell adhesion, membrane ruffling, and microvilli formation (1). ERM proteins undergo intra or intermolecular interaction between their amino- and carboxy-terminal domains, existing as inactive cytosolic monomers or dimers (2). Phosphorylation at a carboxy-terminal threonine residue (Thr567 of ezrin, Thr564 of radixin, Thr558 of moesin) disrupts the amino- and carboxy-terminal association and may play a key role in regulating ERM protein conformation and function (3,4). Phosphorylation at Thr567 of ezrin is required for cytoskeletal rearrangements and oncogene-induced transformation (5). Ezrin is also phosphorylated at tyrosine residues upon growth factor stimulation. Phosphorylation of Tyr353 of ezrin transmits a survival signal during epithelial differentiation (6).

1. Tsukita, S. and Yonemura, S. (1999) J Biol Chem 274, 34507-10.
2. Mangeat, P. et al. (1999) Trends Cell Biol 9, 187-92.
3. Matsui, T. et al. (1998) J Cell Biol 140, 647-57.
4. Gautreau, A. et al. (2000) J Cell Biol 150, 193-203.
5. Tran Quang, C. et al. (2000) EMBO J 19, 4565-76.
6. Gautreau, A. et al. (1999) Proc Natl Acad Sci U S A 96, 7300-5.

注意事项：

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