SignalSilence? UBE2T siRNA I

• 产品编号：CST-13142S      品牌：CST       原厂货号：13142S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® UBE2T siRNA I抑制人、猴中UBE2T的表达。来自Cell Signaling Technology (CST)的SignalSilence^® UBE2T siRNA I可以帮助研究者通过RNA干扰特异性地抑制 UBE2T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^® UBE2T 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。每小瓶可供100次转染，每次转染量相当于在转染24孔板时，每孔总体积为300μl培养基中siRNA的终浓度为100nM。

蛋白泛素化作用需要E1, E2, 和E3泛素-连接酶的协同作用。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-氨基的赖氨酸残疾靶蛋白，这由泛素连接酶E3介导(1)。

UBE2T是E2家族成员，负责ATP依赖性的蛋白质降解。研究发现UBE2T在范科尼贫血中扮演重要角色，并且通过这种途径参与一般的DNA修复过程。UBE2T与FANCL的相互作用激活UBE2T自单泛素化，导致UBE2T的失活和范科尼贫血的负调控（2-4）。另外研究表明， 在乳腺癌和一些肺癌中UBE2T的表达上调，表明其可能参与肿瘤恶化(5,6)。

1. Hershko, A. (1988) J Biol Chem 263, 15237-40.
2. Machida, Y.J. et al. (2006) Mol Cell 23, 589-96.
3. Ramaekers, C.H. et al. (2011) Radiother Oncol 101, 190-7.
4. Zhang, Y. et al. (2007) J Genet Genomics 34, 573-80.
5. Ueki, T. et al. (2009) Cancer Res 69, 8752-60.
6. Hao, J. et al. (2008) Tumour Biol 29, 195-203.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Monkey

Specificity / Sensitivity

SignalSilence^® UBE2T siRNA I inhibits human and monkey UBE2T expression.

Description

SignalSilence^® UBE2T siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit UBE2T 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.

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 ubiquitin from E2 to an ε-amino group of the target protein lysine residue, which is mediated by ubiquitin-ligase enzyme E3 (1).

Ubiquitin conjugating-enzyme 2T (UBE2T) is an E2 family member responsible for the ATP-dependent ubiquitin tagging of target proteins for degradation. Research studies indicate that UBE2T plays an important role in the Fanconi anemia pathway and that UBE2T expression is required for normal DNA repair through this pathway. Interaction between UBE2T and FANCL appears to stimulate UBE2T auto monoubiquitination, leading to UBE2T inactivation and negative regulation of the Fanconi anemia pathway (2-4). Additional research details upregulation of UBE2T expression in breast cancer cells and certain lung carcinomas, suggesting a possible involvement in these malignancies (5,6).

1. Hershko, A. (1988) J Biol Chem 263, 15237-40.
2. Machida, Y.J. et al. (2006) Mol Cell 23, 589-96.
3. Ramaekers, C.H. et al. (2011) Radiother Oncol 101, 190-7.
4. Zhang, Y. et al. (2007) J Genet Genomics 34, 573-80.
5. Ueki, T. et al. (2009) Cancer Res 69, 8752-60.
6. Hao, J. et al. (2008) Tumour Biol 29, 195-203.

注意事项：

Storage: UBE2T siRNA I is supplied in RNAse-free water. Aliquot and store at -20ºC.