SignalSilence? VASP siRNA I

• 产品编号：CST-7637S      品牌：CST       原厂货号：7637S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司的SignalSilence® VASP siRNA I允许研究者通过RNA干扰去特异性抑制VASP蛋白表达，而RNA干扰是通过双链RNA分子传送到细胞中能选择性沉默基因表达的一个方法。所有来自CST公司的SignalSilence® siRNA产品都经过严格的内部检测，且显示通过免疫印迹分析方法减少靶蛋白表达。寡核苷酸的合成是通过三苯基分析（trityl analysis）一个碱基接着一个碱基被监测以确保合适的连接效率。随后通过亲和固相萃取纯化核苷酸。进一步，退火RNA双链通过质谱分析以确保这个双链确切组成。每一批通过质谱分析与前一批比较以确保最大的批间一致。CST公司推荐在细胞裂解前48到72小时转染100 nM SignalSilence® VASP siRNA I。对于转染程序，按照转染试剂商提供的操作步骤。在使用中出现任何问题请随时联系CST公司。Vasodilator-stimulated phosphoprotein (VASP)起初的特征是分别作为cGMP- 和cAMP-依赖的激酶(PKG和PKA或cGPK 和cAPK)的一个底物(1)。研究认为VASP蛋白属于受体蛋白的Ena/VASP家族，而受体蛋白是可将细胞骨架系统联系到信号转导通路，并且其在细胞骨架的组织、成纤维细胞迁移、血小板活化和轴突导向中有一定作用(2,3)。三个磷酸化位点Ser157、Ser239和Thr278已经被鉴定。Ser239位点是主要的PKG磷酸化位点，而Ser157位点是主要的PKA磷酸化位点(4)。研究证明VASP磷酸化可减少它与actin的联系，并且有一个负性影响在actin的聚合作用 (5)。VASP蛋白Ser239位点的磷酸化是对监控PKG激活和信号发送起到一个很好的标记物的作用(6,7)。

1. Butt, E. et al. (1994) J. Biol. Chem. 269, 14509-14517.
2. Ball, L.J. et al. (2000) EMBO J. 19, 4903-4914.
3. Machesky, L.M. et al. (2000) Cell 101, 685-688.
4. Smolenski, A. et al. (1998) J. Biol. Chem. 273, 20029-20035.
5. Harbeck, B. et al. (2000) J. Biol. Chem. 275, 30817-30825.
6. Oelze, M. et al. (2000) Circ. Res. 87, 999-1005.
7. Lawrence, D.W. et al. (2001) J. Immunol. 166, 5550-5556.

原厂资料：

Description

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

Vasodilator-stimulated phosphoprotein (VASP) was originally characterized as a substrate of both cGMP- and cAMP-dependent kinases (PKG and PKA, or cGPK and cAPK, respectively) (1). It is now believed that VASP belongs to the Ena/VASP family of adaptor proteins linking the cytoskeletal system to the signal transduction pathways and that it functions in cytoskeletal organization, fibroblast migration, platelet activation and axon guidance (2,3). Three phosphorylation sites, Ser157, Ser239, and Thr278, have been identified. Ser239 is the major PKG phosphorylation site while Ser157 is the major PKA phosphorylation site (4). Evidence suggests that VASP phosphorylation reduces its association with actin and has a negative effect on actin polymerization (5). Phosphorylation at Ser239 of VASP is a useful marker for monitoring PKG activation and signaling (6,7).

1. Butt, E. et al. (1994) J. Biol. Chem. 269, 14509-14517.
2. Ball, L.J. et al. (2000) EMBO J. 19, 4903-4914.
3. Machesky, L.M. et al. (2000) Cell 101, 685-688.
4. Smolenski, A. et al. (1998) J. Biol. Chem. 273, 20029-20035.
5. Harbeck, B. et al. (2000) J. Biol. Chem. 275, 30817-30825.
6. Oelze, M. et al. (2000) Circ. Res. 87, 999-1005.
7. Lawrence, D.W. et al. (2001) J. Immunol. 166, 5550-5556.

注意事项：

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