SignalSilence? Puma siRNA II

• 产品编号：CST-12890S      品牌：CST       原厂货号：12890S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® Puma siRNA II抑制人和猴Puma表达。来自Cell Signaling Technology (CST)的SignalSilence^® Puma siRNA II 可以帮助研究者通过RNA干扰特异性地抑制Puma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^® Puma siRNA II 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。Puma（P53上调凋亡调节子） ，是一个“BH3-only” Bcl-2家族蛋白，最初确定为一个p53诱导的差异表达基因（1,2） 。 “BH3-only”家族成员包括Bad、Bik、Bid、Bim、Hrk和Noxa，所有这些都包含BH3结构域，但缺乏其他的保守结构域，BH1和BH2 ，通过BH3结构域的相互作用结合或拮抗Bcl-2抗凋亡家族成员，进而促进细胞凋亡（3）。puma基因产生两个含BH3的蛋白质Puma-α和Puma-β，这两者都是由p53诱导产生，结合Bcl-2和Bcl-xL，定位到线粒体，促进细胞色素c的释放和细胞凋亡（1,2）。Puma在p53肿瘤抑制途径中起着至关重要的作用。靶向性地破坏Puma基因，损害p53介导的细胞凋亡和肿瘤抑制（4-7）。研究表明，Puma基因敲除小鼠具有多凋亡刺激缺陷，包括电离辐射、 c-Myc基因表达失调和细胞因子停止（4）。

1. Yu, J. et al. (2001) Mol Cell 7, 673-82.
2. Nakano, K. and Vousden, K.H. (2001) Mol Cell 7, 683-94.
3. Bouillet, P. and Strasser, A. (2002) J Cell Sci 115, 1567-74.
4. Jeffers, J.R. et al. (2003) Cancer Cell 4, 321-8.
5. Hemann, M.T. et al. (2004) Proc Natl Acad Sci U S A 101, 9333-8.
6. Yu, J. et al. (2003) Proc Natl Acad Sci U S A 100, 1931-6.
7. Villunger, A. et al. (2003) Science 302, 1036-8.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® Puma siRNA II inhibits human and monkey Puma expression.

Description

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

Puma (p53 upregulated modulator of apoptosis) is a "BH3-only" Bcl-2 family member originally identified in differential gene expression studies as a p53-inducible gene (1,2). The "BH3-only" family members include Bad, Bid, Bik, Hrk, Bim, and Noxa, all of which contain a BH3 domain but lack other conserved domains, BH1 and BH2, and generally promote apoptosis by binding to and antagonizing anti-apoptotic Bcl-2 family members through BH3 domain interactions (3). Two BH3-containing proteins are produced from the puma gene, Puma-α and Puma-β, both of which are induced by p53, bind Bcl-2 and Bcl-xL, localize to the mitochondria, and promote cyctochrome c release and apoptosis (1,2). Puma plays a critical role in the p53 tumor suppressor pathway. Targeted disruption of the puma gene impairs p53-mediated apoptosis and tumor suprression (4-7). Puma knockout mice show defects from multiple apoptotic stimuli, including ionizing irradiation, deregulated c-Myc expression, and cytokine withdrawal (4).

1. Yu, J. et al. (2001) Mol Cell 7, 673-82.
2. Nakano, K. and Vousden, K.H. (2001) Mol Cell 7, 683-94.
3. Bouillet, P. and Strasser, A. (2002) J Cell Sci 115, 1567-74.
4. Jeffers, J.R. et al. (2003) Cancer Cell 4, 321-8.
5. Hemann, M.T. et al. (2004) Proc Natl Acad Sci U S A 101, 9333-8.
6. Yu, J. et al. (2003) Proc Natl Acad Sci U S A 100, 1931-6.
7. Villunger, A. et al. (2003) Science 302, 1036-8.

注意事项：

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