SignalSilence? PAK1 siRNA II

• 产品编号：CST-6536S      品牌：CST       原厂货号：6536S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

PAK1 siRNA II将抑制人源和大鼠PAK1蛋白表达。Cell Signaling Technology (CST)公司的SignalSilence® PAK1 siRNA II允许研究者通过RNA干扰去特异性抑制PAK1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 PAK1 siRNA II。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。丝氨酸/苏氨酸激酶的p21-activated kinase (PAK)家族是从事于多种细胞内过程，包括细胞骨架的重排、MAPK信号、凋亡信号、吞噬细胞NADPH氧化酶的控制和生长因子诱导的轴突生长(1,2)。诱导PAK激活的数种机制已经被报道。Rac/Cdc42结合到PAK的氨基端CRIB (or PBD)结构域上引起PAK的自磷酸和构象的改变(1)。通过 PDK使PAK1在Thr423位点磷酸化诱导PAK1的激活(3)。数个自磷酸化位点已经被鉴定，包括PAK1的Ser199和Ser204位点以及PAK2的Ser192和Ser197位点(4,5)。因为自磷酸化位点位于氨基端抑制结构域，所以推测在这个区域内的修饰抑制该激酶恢复到失活构象(6)。研究指出PAK1的Ser144位点磷酸化或PAK3的Ser139位点(位于该激酶抑制区域)影响激酶活性(7)。PAK1的Ser21位点或PAK2的Ser20位点的磷酸化调节与接头蛋白Nck的结合(8)。PAK4、PAK5和PAK6与PAK1-3在氨基端调节域有低的相同序列(9)。PAK4的Ser474位点的磷酸化类似于PAK1的Thr423位点，它可能在调节PAK4的活性和功能起着重要作用(10)。

1. Knaus, U.G. and Bokoch, G.M. (1998) Int. J. Biochem. Cell Biol. 30, 857-862.
2. Daniels, R.H. et al. (1998) EMBO J. 17, 754-764.
3. King, C.C. et al. (2000) J. Biol. Chem. 275, 41201-41209.
4. Manser, E. et al. (1997) Mol. Cell. Biol. 17, 1129-1143.
5. Gatti, A. et al. (1999) J. Biol. Chem. 274, 8022-8028.
6. Lei, M. et al. (2000) Cell 102, 387-397.
7. Chong, C. et al. (2001) J. Biol. Chem. 276, 17347-17353.
8. Zhao, Z. et al. (2000) Mol. Cell. Biol. 20, 3906-3917.
9. Abo, A. et al. (1998) EMBO J. 17, 6527-6540.
10. Qu, J. et al. (2001) Mol. Cell. Biol. 21, 3523-3533.

原厂资料：

Homology

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

Specificity / Sensitivity

PAK1 siRNA II will inhibit human and rat PAK1 expression.

Description

SignalSilence® PAK1 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit PAK1 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 are rigorously tested in-house and have been shown to reduce 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 PAK1 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

The p21-activated kinase (PAK) family of serine/threonine kinases is engaged in multiple cellular processes, including cytoskeletal reorganization, MAPK signaling, apoptotic signaling, control of phagocyte NADPH oxidase, and growth factor-induced neurite outgrowth (1,2). Several mechanisms that induce PAK activity have been reported. Binding of Rac/Cdc42 to the CRIB (or PBD) domain near the amino terminus of PAK causes autophosphorylation and conformational changes in PAK (1). Phosphorylation of PAK1 at Thr423 by PDK induces activation of PAK1 (3). Several autophosphorylation sites have been identified, including Ser199 and Ser204 of PAK1 and Ser192 and Ser197 of PAK2 (4,5). Because the autophosphorylation sites are located in the amino-terminal inhibitory domain, it has been hypothesized that modification in this region prevents the kinase from reverting to an inactive conformation (6). Research indicates that phosphorylation at Ser144 of PAK1 or Ser139 of PAK3 (located in the kinase inhibitory domain) affects kinase activity (7). Phosphorylation at Ser21 of PAK1 or Ser20 of PAK2 regulates binding with the adaptor protein Nck (8). PAK4, PAK5, and PAK6 have lower sequence similarity with PAK1-3 in the amino-terminal regulatory region (9). Phosphorylation at Ser474 of PAK4, a site analogous to Thr423 of PAK1, may play a pivotal role in regulating the activity and function of PAK4 (10).

1. Knaus, U.G. and Bokoch, G.M. (1998) Int. J. Biochem. Cell Biol. 30, 857-862.
2. Daniels, R.H. et al. (1998) EMBO J. 17, 754-764.
3. King, C.C. et al. (2000) J. Biol. Chem. 275, 41201-41209.
4. Manser, E. et al. (1997) Mol. Cell. Biol. 17, 1129-1143.
5. Gatti, A. et al. (1999) J. Biol. Chem. 274, 8022-8028.
6. Lei, M. et al. (2000) Cell 102, 387-397.
7. Chong, C. et al. (2001) J. Biol. Chem. 276, 17347-17353.
8. Zhao, Z. et al. (2000) Mol. Cell. Biol. 20, 3906-3917.
9. Abo, A. et al. (1998) EMBO J. 17, 6527-6540.
10. Qu, J. et al. (2001) Mol. Cell. Biol. 21, 3523-3533.

注意事项：

Storage: PAK1 siRNA II is supplied in RNAse-free water. Aliquot
and store at -20°C.