SignalSilence? Apaf-1 siRNA I

• 产品编号：CST-6229S      品牌：CST       原厂货号：6229S
• 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^® Apaf-1 siRNA I 可以帮助研究者通过RNA干扰特异性地抑制Apaf-1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^® Apaf-1 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。凋亡蛋白酶激活因子 1 (Apaf-1)是凋亡过程中激活caspase-9的一个重要信号蛋白，人们最初把它鉴定为哺乳动物中与C. elegans 凋亡调节蛋白CED-4同源的蛋白(1)。在细胞色素C与dATP存在下，凋亡因子Apaf-1与caspase-9形成复合体，极大的促进了caspase-9的激活，进而导致caspase-3的激活 (2,3)。此蛋白包含一个N末端的CARD域，一个中间的CED-4同源域和C末端的多个WD-40重复。通过在CARD结构域之后的小片段插入一个多余的WD-40重复 (4)等可变剪切，Apaf-1能够表达多种亚型。Apaf-1敲除小鼠模型的研究表明，其存在普遍的凋亡缺陷并且能够抵抗很多凋亡刺激(5,6)。

1. Zou, H. et al. (1997) Cell 90, 405-13.
2. Zou, H. et al. (1999) J Biol Chem 274, 11549-56.
3. Saleh, A. et al. (1999) J Biol Chem 274, 17941-5.
4. Benedict, M.A. et al. (2000) J Biol Chem 275, 8461-8.
5. Cecconi, F. et al. (1998) Cell 94, 727-37.
6. Yoshida, H. et al. (1998) Cell 94, 739-50.

原厂资料：

Description

SignalSilence® Apaf-1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Apaf-1 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 Apaf-1 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

Apoptotic protease activating factor 1 (Apaf-1), originally identified as the mammalian homolog of the C. elegans apoptotic regulatory protein CED-4, is an important signaling protein involved in the activation of caspase-9 during apoptosis (1). Cytosolic Apaf-1 forms a complex with caspase-9 in the presence of cytochrome c and dATP, ultimately leading to caspase-9 activation and subsequent activation of caspase-3 (2,3). The protein contains an amino-terminal CARD domain, a central CED-4 homology domain, and multiple WD-40 repeats at the carboxy-terminus. Several isoforms of Apaf-1 are expressed through alternative splicing generating a small insert following the CARD domain as well as an extra WD-40 repeat (4). Apaf-1 knock-out mice display widespread defects in apoptosis and resistance to a variety of apoptotic stimuli (5,6).

1. Zou, H. et al. (1997) Cell 90, 405-13.
2. Zou, H. et al. (1999) J Biol Chem 274, 11549-56.
3. Saleh, A. et al. (1999) J Biol Chem 274, 17941-5.
4. Benedict, M.A. et al. (2000) J Biol Chem 275, 8461-8.
5. Cecconi, F. et al. (1998) Cell 94, 727-37.
6. Yoshida, H. et al. (1998) Cell 94, 739-50.

注意事项：

Storage: Apaf-1 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.