SignalSilence? UBE2S siRNA II

• 产品编号：CST-7732S      品牌：CST       原厂货号：7732S
• 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^®UBE2S siRNA II可以帮助研究者通过RNA干扰特异性地抑制UBE2S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^® UBE2S siRNA II 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。蛋白泛素化作用需要E1, E2, 和E3泛素-结合酶的协同作用。泛素最先通过具有硫酯的泛素激活酶E1通过ATP-依赖形式活化。活化的泛素然后通过含有巯基的泛素载体蛋白E2转运。最后从E2的泛素转运到含有ε-氨基的赖氨酸残基靶蛋白，这由泛素连接酶E3介导(1)。人分裂后期促进复合物(APC/C)是一种大分子的E3连接酶复合物，经过对蛋白酶体降解的细胞周期的调节子的连续的靶向作用，对有丝分裂的时序性进程起重要作用。最近的研究表明APC/C的底物通过共价组装Lys11-连接的泛素链，在细胞周期进展中起蛋白酶体的降解作用，主要发生在起始阶段和延伸阶段(2-5)。APC/C部分利用UBE2C/UBCH10 E2酶来介导底物的降解，通过与短的Lys11-linked链共价连接(3,6)。Lys11-特异的延伸的E2酶，UBE2S/E2-EPF, 扩展这些短链到APC/C 结合底物上的Lys11-linked泛素链(2,3,7)。除外在细胞周期调节中的UBE2S的生化作用被明确外，这些酶在许多人癌症中过度表达(8)，并在缺氧信号传导中发挥作用(9,10)。确实，UBE2S已经被报道与VHL有关，并是蛋白酶体降解的靶目标，从而稳定了HIF-1α (9)。

1. Hershko, A. (1988) J Biol Chem 263, 15237-40.
2. Williamson, A. et al. (2009) Proc Natl Acad Sci USA 106, 18213-8.
3. Jin, L. et al. (2008) Cell 133, 653-65.
4. Wu, T. et al. (2010) Proc Natl Acad Sci USA 107, 1355-60.
5. Song, L. and Rape, M. (2010) Mol Cell 38, 369-82.
6. Summers, M.K. et al. (2008) Mol Cell 31, 544-56.
7. Wickliffe, K.E. et al. (2011) Cell 144, 769-81.
8. Tedesco, D. et al. (2007) Neoplasia 9, 601-13.
9. Jung, C.R. et al. (2006) Nat Med 12, 809-16.

原厂资料：

Description

SignalSilence® UBE2S siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit UBE2S 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 SignalSilence® UBE2S 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

Protein ubiquitination requires the concerted action of the E1, E2, and E3 ubiquitin-conjugating enzymes. Ubiquitin is first activated through ATP-dependent formation of a thiol ester with ubiquitin-activating enzyme E1. The activated ubiquitin is then transferred to a thiol group of ubiquitin-carrier enzyme E2. The final step is the transfer of ubiqThe human anaphase promoting complex (APC/C) is a large macromolecular E3 ligase complex that is largely responsible for timely progression through mitosis via the sequential targeting of cell cycle regulators for proteasomal degradation. Recent work has revealed that APC/C substrates are marked for proteasomal degradation during cell cycle progression through the covalent assembly of Lys11-linked ubiquitin chains, which occurs through a priming phase and an elongation phase (2-5). The APC/C utilizes, in part, the UBE2C/UBCH10 E2 enzyme to prime substrates for degradation through the covalent attachment of short Lys11-linked chains (3,6). The Lys11-specific elongating E2 enzyme, UBE2S/E2-EPF, extends these short chains into long Lys11-linked ubiquitin chains on APC/C bound substrates (2,3,7). In addition to the well-established biochemical role for UBE2S in cell cycle regulation, researchers have found evidence that this enzyme is overexpressed in many types of human cancer (8), and has been implicated in hypoxia signaling (9,10). Indeed, UBE2S has been reported by researchers to associate with VHL and to target it for proteasomal degradation, thereby stabilizing HIF-1α (9).

1. Hershko, A. (1988) J Biol Chem 263, 15237-40.
2. Williamson, A. et al. (2009) Proc Natl Acad Sci USA 106, 18213-8.
3. Jin, L. et al. (2008) Cell 133, 653-65.
4. Wu, T. et al. (2010) Proc Natl Acad Sci USA 107, 1355-60.
5. Song, L. and Rape, M. (2010) Mol Cell 38, 369-82.
6. Summers, M.K. et al. (2008) Mol Cell 31, 544-56.
7. Wickliffe, K.E. et al. (2011) Cell 144, 769-81.
8. Tedesco, D. et al. (2007) Neoplasia 9, 601-13.
9. Jung, C.R. et al. (2006) Nat Med 12, 809-16.

注意事项：

Storage: UBE2S siRNA II is supplied in RNAse-free water.
Aliquot and store at -20ºC