SignalSilence? NDP52 siRNA I

• 产品编号：CST-8964S      品牌：CST       原厂货号：8964S
• 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^® NDP52 siRNA I可以帮助研究者通过RNA干扰特异性地抑制NDP52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^® NDP52 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。NDP52 (CALCOCO2) 广泛表达，由N-末端的SKICH域、卷曲螺旋域和两个锌指域构成(1)。最近的研究表明它作为一个自噬受体，能够结合细胞质中带有泛素化修饰的细菌，导致自噬的激活和病原体的清除(1-3)。NDP52 通过其锌指域结合泛素，同时结合到LC3并指引细菌进入自噬小体。另外，NDP52与Nap1和SINTBAD相互作用，将TBK1招募到泛素化的细菌上(1)。

1. Thurston, T.L. et al. (2009) Nat Immunol 10, 1215-21.
2. Cemma, M. et al. (2011) Autophagy 7, 341-5.
3. Mostowy, S. et al. (2011) J Biol Chem 286, 26987-95.

原厂资料：

Homology

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

Description

SignalSilence® NDP52 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit NDP52 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® NDP52 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. Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

NDP52 (CALCOCO2) is ubiquitously expressed and composed of an amino-terminal SKICH domain, followed by a coiled-coil domain, and two zinc finger domains (1). It has recently been reported to act as an autophagy receptor that binds cytosolic ubiquitinated bacteria, leading to autophagy activation and pathogen clearance (1-3). NDP52 binds ubiquitin through its zinc finger domain and simultaneously binds LC3, which directs the bacteria into autophagosomes. In addition, NDP52 interacts with Nap1 and SINTBAD to recruit TBK1 to ubiquitinated bacteria (1).

1. Thurston, T.L. et al. (2009) Nat Immunol 10, 1215-21.
2. Cemma, M. et al. (2011) Autophagy 7, 341-5.
3. Mostowy, S. et al. (2011) J Biol Chem 286, 26987-95.

注意事项：

Storage: NDP52 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC