SignalSilence? RKIP siRNA II

• 产品编号：CST-6297S      品牌：CST       原厂货号：6297S
• 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^® RKIP siRNA II可以帮助研究者通过RNA干扰特异性地抑制RKIP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^® RKIP 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。Raf 激酶抑制蛋白(RKIP)是磷脂酰乙醇胺结合蛋白(PEBP)家族成员，该家族与Raf-1、 MEK、MAPK相关(1)。RKIP已被证明能与Raf-1, MEK, Erk形成复合物 (2)。尽管MEK、Erk能同时结合RKIP，但是Raf-1与RKIP、RKIP与MEK之间的结合是相互排斥的。因此，RKIP可以竞争性地破坏Raf-1-MEK复合物，并有效地终止从Raf-1到MAPK的信号传递(2)。RKIP对MAPK信号转导的抑制效应，会因为RKIP的Ser153位点被PKC磷酸化而得到消除(3)。PKC对RKIP的Ser153位点的磷酸化也促进了RKIP与GRK2的结合，从而阻止了GRK2依赖的GPCR的内化(4)。RKIP也能与NF-κB通路中的分子相互作用，包括NF-κB诱导的激酶(NIK), TAK1, IKKα ，IKKβ (5)。这些相互作用能够对细胞因子诱导的NF-κB通路的活化起到拮抗作用(5)。RKIP表达的恢复与前列腺癌症转移的抑制有关，暗示RKIP可能是肿瘤转移的抑制子从而成为潜在的临床靶点，这种抑制作用是通过抑制血管侵润而实现的(6)。

1. Yeung, K. et al. (1999) Nature 401, 173-7.
2. Yeung, K. et al. (2000) Mol Cell Biol 20, 3079-85.
3. Corbit, K.C. et al. (2003) J Biol Chem 278, 13061-8.
4. Lorenz, K. et al. (2003) Nature 426, 574-9.
5. Yeung, K.C. et al. (2001) Mol Cell Biol 21, 7207-17.
6. Fu, Z. et al. (2003) J Natl Cancer Inst 95, 878-89.

原厂资料：

Description

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

Background

Raf kinase inhibitor protein (RKIP) is a member of the phosphatidylethanolamine-binding protein (PEBP) family that associates with Raf-1 and the MEK and MAP kinases (1). RKIP has been shown to complex with Raf-1, MEK, and ERK (2). Although MEK and ERK can simultaneously bind RKIP, the association between Raf-1 and RKIP and that of RKIP and MEK are mutually exclusive. Thus, RKIP competitively disrupts the Raf-1-MEK complex and effectively terminates signal transmission from Raf-1 to MAP kinases (2). The inhibitory effect of RKIP on MAP kinase signaling is eliminated by PKC phosphorylation of RKIP at Ser153 (3). PKC phosphorylation on Ser153 also promotes the association of RKIP with GRK2, which prevents GRK2-dependent internalization of GPCR (4). RKIP also interacts with modules of the NF-κB pathway, including NF-κB-inducing kinase (NIK), TAK1, IKKα and IKKβ (5). These interactions antagonize cytokine-induced activation of the NF-κB pathway (5). Restoration of RKIP expression is associated with the inhibition of prostate cancer metastasis, implying that RKIP may be a potential clinical target as a suppressor of tumor metastasis through inhibition of vascular invasion (6).

1. Yeung, K. et al. (1999) Nature 401, 173-7.
2. Yeung, K. et al. (2000) Mol Cell Biol 20, 3079-85.
3. Corbit, K.C. et al. (2003) J Biol Chem 278, 13061-8.
4. Lorenz, K. et al. (2003) Nature 426, 574-9.
5. Yeung, K.C. et al. (2001) Mol Cell Biol 21, 7207-17.
6. Fu, Z. et al. (2003) J Natl Cancer Inst 95, 878-89.

注意事项：

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