SignalSilence? FLIP siRNA II

• 产品编号：CST-12407S      品牌：CST       原厂货号：12407S
• 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^® FLIP siRNA II 可以帮助研究者通过RNA干扰特异性地抑制FLIP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^® FLIP 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。细胞的FLIP（FLICE抑制蛋白）是一种细胞凋亡调节子，其有很多种名称，如c-FLIP (1)、Casper (2)、CLARP (3)、FLAME (4)、I-FLICE (5)、MRIT (6)、CASH (7)和Usurpin (8)。FLIP以两种剪接形式选择性表达， 即短FLIP（FLIPS）和长FLIP（FLIPL）。FLIPS包含两个死亡效应域（DEDS），发现存在于死亡受体衔接蛋白FADD和caspase-8前体域。FLIPL与caspase-8（FLICE）共享重要的同源域，并包含一个额外的死亡效应域，但FLIPL缺乏半胱天冬酶的催化活性位点，并且不具有蛋白酶活性。据研究报道，这两种形式都与FADD和caspase-8前体相互作用。关于FLIP在凋亡中的作用的研究是有争议性的，一些报道是抗细胞凋亡，而另一些则是促凋亡。FLIPL的过表达可导致caspase-8异源化从而产生活性蛋白酶，进而导致细胞凋亡。然而在生理水平，研究认为FLIP结合到FADD的DED会抑制caspase-8。通过siRNA或基因打靶降低FLIP表达，会使细胞对死亡受体介导的细胞凋亡敏感。FLIP也与癌细胞凋亡的阻滞相关，在某些类型的癌症包括Hodgkin's 淋巴瘤、卵巢癌和结肠癌中上调表达（9）。

1. Irmler, M. et al. (1997) Nature 388, 190-195.
2. Shu, H. B. et al. (1997) Immunity 6, 751-763.
3. Inohara, N. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 10717-10722.
4. Srinivasula, S. M. et al. (1997) J. Biol. Chem. 272, 18542-18545.
5. Hu, S. et al. (1997) J. Biol. Chem. 272, 17255-17257.
6. Han, D. K. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 11333-11338.
7. Dita, R. M. et al. (1998) Cell Death Differ. 5, 271-288.
8. Goltsev, Y. V. et al. (1997) J. Biol. Chem. 272, 19641-19644.
9. Kataoka, T. (2005) Crit. Rev. Immunol. 25, 31-58.

原厂资料：

Description

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

Cellular FLIP (FLICE inhibitory protein) is a regulator of apoptosis that has various names, such as c-FLIP (1), Casper (2), CLARP (3), FLAME (4), I-FLICE (5), MRIT (6), CASH (7), and Usurpin (8). FLIP is expressed as two alternative splice isoforms, FLIP short (FLIPS) and FLIP long (FLIPL). FLIPS contains two death effector domains (DEDs) like those found on the death receptor adaptor protein FADD and the pro-domain of caspase-8. FLIPL shares significant homology with caspase-8 (FLICE), and contains an additional death effector domain, but FLIPL lacks the catalytic active site of the caspases and does not have protease activity. Both FLIP isoforms have been reported to interact with FADD and pro-caspase-8. The role of FLIP in apoptosis is controversial as some research studies have reported it to be anti-apoptotic, while others claim that it is pro-apoptotic. Overexpression of FLIPL can lead to caspase-8 heterodimers that produce an active protease, resulting in apoptosis. However, at physiological levels, it is thought that the binding of FLIP to the DED of FADD results in inhibition of caspase-8 processing. Reduction of FLIP by siRNA or gene targeting sensitizes cells to death receptor-mediated apoptosis. FLIP has also been implicated in the resistance of cancer cells to apoptosis and is upregulated in some cancer types including Hodgkin's lymphoma and ovarian and colon carcinomas (9).

1. Irmler, M. et al. (1997) Nature 388, 190-195.
2. Shu, H. B. et al. (1997) Immunity 6, 751-763.
3. Inohara, N. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 10717-10722.
4. Srinivasula, S. M. et al. (1997) J. Biol. Chem. 272, 18542-18545.
5. Hu, S. et al. (1997) J. Biol. Chem. 272, 17255-17257.
6. Han, D. K. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 11333-11338.
7. Dita, R. M. et al. (1998) Cell Death Differ. 5, 271-288.
8. Goltsev, Y. V. et al. (1997) J. Biol. Chem. 272, 19641-19644.
9. Kataoka, T. (2005) Crit. Rev. Immunol. 25, 31-58.

注意事项：

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