Cell Signaling Technology

Product Pathways - Protein Stability

SignalSilence® USP9X siRNA I #6308

ubiquitin  

No. Size Price
6308S 300 µl ( 3 nmol ) ¥3,224.00 现货查询 购买询价
6308 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
TFN Human,

Species cross-reactivity is determined by western blot.

Applications Key: TFN=Transfection,

Source / Purification

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® USP9X siRNA I (+), using USP9X Antibody #5751 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The USP9X Antibody confirms silencing of USP9X expression, while theα-Tubulin (11H10) Rabbit mAb is used as a loading control.

使用USP9X Antibody #5751 (上)或α-Tubulin (11H10) Rabbit mAb #2125(下)对分别转染100nM SignalSilence® 对照siRNA(无偶联物)#6568(-)、SignalSilence® USP9X siRNA I (+)的HeLa细胞提取物进行免疫印迹分析结果。USP9X#5751抗体证实USP9X的表达被沉默,而α-Tubulin (11H10) Rabbit mAb #2125抗体用于上样对照。

Description

来自Cell Signaling Technology (CST)的SignalSilence® USP9X siRNA可以帮助研究者通过RNA干扰特异性地抑制USP9X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。

Quality Control

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。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® USP9X siRNA I (+), using USP9X Antibody #5751 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The USP9X Antibody confirms silencing of USP9X expression, while theα-Tubulin (11H10) Rabbit mAb is used as a loading control.

Directions for Use

CST推荐使用100 nM SignalSilence® USP9X 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。

Background

泛素酶(UBEs)催化蛋白质泛素化,去泛素化酶(DUB)作用相反(1,2)。共有5个亚家族的去泛素化酶被发现,包括USP, UCH, OTU, MJD和JAMM酶。USP9X/hFAM是一个X染色体连锁的泛素特异性蛋白酶亚家族去泛素化酶成员,含有一个非常保守的催化结构域,具有半胱氨酸肽酶活性,使其能够对泛素或者多泛素偶联分子进行切割。USP9X是果蝇fat-facets (faf)基因在 哺乳动物中的同源基因,而faf基因对于果蝇眼部发育和胚胎存活能力至关重要(3,4)。 尽管USP9X对于哺乳动物胚胎发育也非常重要(5-7),但其底物分子的鉴定才刚刚开始。有多项证据表明,USP9X在表皮细胞极化过程中,通过去泛素化活性依赖的方式,提高粘附分子和紧密连接蛋白的稳定性,从而参与细胞与细胞之间接触形成。实际上,USP9X被发现与AF-6、β-catenin-E-cadherin复合体及EFA6有相互作用(8-11)。也有研究发现,USP9X是TGF-β/BMP信号级联途径不可或缺的成员,能够抑制TRIM33介导的SMAD4的单泛素化,从而调节TGF-β的响应(12)。最近,USP9X被发现在多种肿瘤中高表达,通过其去泛素化能力增加Mcl-1蛋白的稳定性, 使癌细胞的生存能力和抗化疗能力大大增加。这一证据提示,USP9X能够在滤泡性淋巴瘤和弥漫性大B细胞淋巴瘤等恶性肿瘤中作为治疗对象(13)。

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalSilence is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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