SignalSilence? Atg5 siRNA I

• 产品编号：CST-6345S      品牌：CST       原厂货号：6345S
• 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^® Atg5 siRNA I 可以帮助研究者通过RNA干扰特异性地抑制Atg5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^® Atg5 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。自噬是自噬溶酶体降解其所包裹的细胞质内容物的一个分解代谢过程(1,2)。通常情况下，自噬在营养匮乏的时候被激活，但它还与许多生理过程相关，包括生长、分化、神经退行性疾病、感染和癌症(3)。自噬的分子机制主要在酵母中发现的，并称之为自噬相关基因。自噬体的形成涉及一个泛素样连接体系，其中Atg12和Atg5共价结合，并定位到自噬体囊泡中(4-6)。上述连接反应由泛素E1样连接酶Atg7和E2样连接酶Atg10介导(7,8)。

1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
3. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
4. Mizushima, N. et al. (1998) J Biol Chem 273, 33889-92.
5. Mizushima, N. et al. (1998) Nature 395, 395-8.
6. Suzuki, K. et al. (2001) EMBO J 20, 5971-81.
7. Tanida, I. et al. (1999) Mol Biol Cell 10, 1367-79.
8. Shintani, T. et al. (1999) EMBO J 18, 5234-41.

原厂资料：

Description

SignalSilence® Atg5 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Atg5 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 Atg5 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

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegeneration, infection, and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes. Formation of the autophagosome involves a ubiquitin-like conjugation system in which Atg12 is covalently bound to Atg5 and targeted to autophagosome vesicles (4-6). This conjugation reaction is mediated by the ubiquitin E1-like enzyme Atg7 and the E2-like enzyme Atg10 (7,8).

1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
3. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
4. Mizushima, N. et al. (1998) J Biol Chem 273, 33889-92.
5. Mizushima, N. et al. (1998) Nature 395, 395-8.
6. Suzuki, K. et al. (2001) EMBO J 20, 5971-81.
7. Tanida, I. et al. (1999) Mol Biol Cell 10, 1367-79.
8. Shintani, T. et al. (1999) EMBO J 18, 5234-41.

注意事项：

Storage: Atg5 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.