| 货号 | 6361S |
| 描述 | SignalSilence® PAK1 siRNA I 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 from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis. |
| 反应种属 | Human |
| 应用 | TFN |
| 目标/特异性 | PAK1 siRNA I will inhibit human and rat PAK1 expression. |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | -20C |
| 计算分子量 | 68 |
| 参考文献 | Knaus, U.G. and Bokoch, G.M. (1998) Int. J. Biochem. Cell Biol. 30, 857-862. Daniels, R.H. et al. (1998) EMBO J. 17, 754-764. King, C.C. et al. (2000) J. Biol. Chem. 275, 41201-41209. Manser, E. et al. (1997) Mol. Cell. Biol. 17, 1129-1143. Gatti, A. et al. (1999) J. Biol. Chem. 274, 8022-8028. Lei, M. et al. (2000) Cell 102, 387-397. Chong, C. et al. (2001) J. Biol. Chem. 276, 17347-17353. Zhao, Z. et al. (2000) Mol. Cell. Biol. 20, 3906-3917. Abo, A. et al. (1998) EMBO J. 17, 6527-6540. Qu, J. et al. (2001) Mol. Cell. Biol. 21, 3523-3533. Wang, R. et al. (2002) EMBO J. 21, 5437-5447. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® PAK1 siRNA I (+), using PAK1 Antibody #2602 and p42 MAP Kinase (Erk2) Antibody #9108. The PAK1 antibody confirms silencing of PAK1 expression, while the p42 MAP kinase antibody is used to control for loading and specificity of PAK1 siRNA. 使用PAK1 Antibody #2602和p42 MAP Kinase (Erk2) Antibody #9108，免疫印迹（Western blot）分析HeLa细胞中PAK1蛋白水平，细胞分别转染100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-)或SignalSilence® PAK1 siRNA I (+)。PAK1 antibody确定PAK1基因沉默，然而p42 MAP kinase antibody通常用于内参照和PAK1 siRNA的特异性。 | |
Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency. 在转染后24小时后的HeLa细胞中SignalSilence® Control siRNA (Fluorescein Conjugate) #6201的荧光检测证明接近100%的转染效率。 |
