Cell Signaling Technology

Product Pathways - Protein Stability

SignalSilence® UBA2 siRNA I #7646

No. Size Price
7646S 300 µl ( 3 nmol ) ¥3,224.00 现货查询 购买询价
7646 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 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® UBA2 siRNA I (+) or SignalSilence® UBA2 siRNA II #7642 (+), using UBA2 (D15C11) Rabbit mAb #8688 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The UBA2 (D15C11) Rabbit mAb confirms silencing of UBA2 expression, while the GAPDH (D16H11) XP® Rabbit mAb is used as a loading control. 使用UBA2 (D15C11)兔单抗 #8688 (上) or GAPDH (D16H11) XP® 兔单抗#5174 (下)对分别转染100nM SignalSilence® 对照siRNA(无偶联物)#6568(-)、 SignalSilence® UBA2 siRNA I (+)或SignalSilence® UBA2 siRNA II #7642 (+)的HeLa细胞提取物进行免疫印迹分析结果。UBA2 (D15C11)兔单抗证实UBA2的表达沉默,而GAPDH (D16H11) XP® 兔单抗#5174 用做上样对照。

Description

来自Cell Signaling Technology (CST)的SignalSilence® UBA2 siRNA I可以帮助研究者通过RNA干扰特异性地抑制UBA2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 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® UBA2 siRNA I (+) or SignalSilence® UBA2 siRNA II #7642 (+), using UBA2 (D15C11) Rabbit mAb #8688 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The UBA2 (D15C11) Rabbit mAb confirms silencing of UBA2 expression, while the GAPDH (D16H11) XP® Rabbit mAb is used as a loading control.

Directions for Use

CST推荐使用100 nM SignalSilence® UBA2 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。

Background

SUMO修饰结合于目标蛋白质的过程是类似于泛素化修饰的事件(1)。SUMO是通过由E1,E2和E3酶(2)组成的SUMO修饰系统协同作用而连接到靶向蛋白。典型的SUMOE1激活酶是由SAE1(AOS1)和UBA2(SAE2)亚基组成的异源二聚体。成熟SUMO是由激活酶E1通过依赖于ATP的反应,生成腺苷酸化SUMO(SUMO与UBA2的Cys173之间形成硫酯键而形成的一种高能中间体)(3,4)。SUMO然后从UBA2转移到SUMO的E2结合酶UBC9(5)。最近的证据表明,UBA2氧化还原调控作为一个生理机制,调节sumo化靶蛋白的细胞水平(6)。

  1. Geiss-Friedlander, R. and Melchior, F. (2007) Nat Rev Mol Cell Biol 8, 947-56.
  2. Tatham, M.H. et al. (2003) Biochemistry 42, 9959-69.
  3. Desterro, J.M. et al. (1999) J Biol Chem 274, 10618-24.
  4. Gong, L. et al. (1999) FEBS Lett 448, 185-9.
  5. Desterro, J.M. et al. (1997) FEBS Lett 417, 297-300.
  6. Bossis, G. and Melchior, F. (2006) Mol Cell 21, 349-57.

Application References

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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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