SignalSilence? Rabex-5 siRNA I

• 产品编号：CST-6437S      品牌：CST       原厂货号：6437S
• 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)公司生产的SignalSilence Rabex-5 siRNA I使得研究者可以使用RNA干扰技术特异性的抑制Rabex-5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公司推荐您在细胞裂解前用100nM Rabex-5 siRNA I进行48-72小时的转染。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。Rabex-5也称为RabGEF1和RAP1，被确定为Rab5的鸟嘌呤核苷酸交换因子（GEF），是小Rab GTP酶Ras超家族的成员之一（1）。Rabex-5从产生GTP结合的活性形式的Rab5，并与其效应蛋白Rabaptin-5形成紧密连接（2）。这个复合物定位到内体膜，作为早期内吞中囊泡转运的关键调节者发挥作用（3,4）。Rabex-5也可以被单泛素化，并且有泛素连接酶活性以便被招募到早期内体（5,6）。Rab5在GTP/GDP状态下的构象改变对其生物学功能至关重要，因为其是内吞作用多个步骤中的一个速率限制调节者（5）。通过其对内体转运和入胞的控制，Rabex-5显示出对NGF介导的神经突生长和FcεRI依赖的肥大细胞活化的负向调节（8）。

1. Horiuchi, H. et al. (1997) Cell 90, 1149-59.
2. Lippé, R. et al. (2001) Mol Biol Cell 12, 2219-28.
3. Zerial, M. and McBride, H. (2001) Nat Rev Mol Cell Biol 2, 107-17.
4. van der Bliek, A.M. (2005) Nat Cell Biol 7, 548-50.
5. Mattera, R. et al. (2006) J Biol Chem 281, 6874-83.
6. Mattera, R. and Bonifacino, J.S. (2008) EMBO J 27, 2484-94.
7. Liu, J. et al. (2007) Mol Biol Cell 18, 1375-84.
8. Tam, S.Y. et al. (2004) Nat Immunol 5, 844-52.

原厂资料：

Description

SignalSilence® Rabex-5 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Rabex-5 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 Rabex-5 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

Rabex-5, also called RabGEF1 and RAP1, was identified as a guanine nucleotide exchange factor (GEF) for Rab5, a member of the Ras superfamily of small Rab GTPases (1). Rabex-5 generates the GTP-bound active form of Rab5 and forms a tight association with its effector protein Rabaptin-5 (2). This complex localizes to endosomal membranes where it functions as a key regulator of vesicular trafficking during early endocytosis (3,4). Rabex-5 is also monoubiquitinated and has ubiquitin ligase activity that regulates its recruitment to early endosomes (5,6). The conformational change between Rab5 GTP/GDP states is essential for its biological function as a rate limiting regulator at multiple steps during endocytosis (5). Through its control of endosomal trafficking and endocytosis, Rabex-5 has been shown to negatively regulate NGF-mediated neurite outgrowth (7) as well as FcεRI-dependent mast cell activation (8).

1. Horiuchi, H. et al. (1997) Cell 90, 1149-59.
2. Lippé, R. et al. (2001) Mol Biol Cell 12, 2219-28.
3. Zerial, M. and McBride, H. (2001) Nat Rev Mol Cell Biol 2, 107-17.
4. van der Bliek, A.M. (2005) Nat Cell Biol 7, 548-50.
5. Mattera, R. et al. (2006) J Biol Chem 281, 6874-83.
6. Mattera, R. and Bonifacino, J.S. (2008) EMBO J 27, 2484-94.
7. Liu, J. et al. (2007) Mol Biol Cell 18, 1375-84.
8. Tam, S.Y. et al. (2004) Nat Immunol 5, 844-52.

注意事项：

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