| 货号 | 6566S |
| 描述 | SignalSilence® p70/85 S6 Kinase siRNA I allows the researcher to specifically inhibit p70/85 S6 kinase 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 target protein expression by western analysis. |
| 反应种属 | Human |
| 应用 | TFN |
| 目标/特异性 | p70/85 S6 Kinase siRNA I will inhibit human, rat and monkey p70/85 S6 Kinase expression. |
| 供应商 | CST |
| 背景 | p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5 oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8). |
| 存放说明 | -20C |
| 参考文献 | Pullen, N. and Thomas, G. (1997) FEBS Lett 410, 78-82. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9. Weng, Q.P. et al. (1998) J Biol Chem 273, 16621-9. Pullen, N. et al. (1998) Science 279, 707-10. Alessi, D.R. et al. (1998) Curr Biol 8, 69-81. Polakiewicz, R.D. et al. (1998) J Biol Chem 273, 23534-41. Fingar, D.C. et al. (2002) Genes Dev 16, 1472-87. Saitoh, M. et al. (2002) J Biol Chem 277, 20104-12. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® p70/85 S6 Kinase siRNA I (+) or SignalSilence® p70/85 S6 Kinase siRNA II #6572 (+), using p70 S6 Kinase (49D7) Rabbit mAb #2708 and α-Tubulin (11H10) Rabbit mAb #2125. The p70 S6 Kinase (49D7) Rabbit mAb confirms silencing of p70/85 S6 kinase expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p70/85 S6 kinase siRNA. Western blot 分析Hela细胞提取物，采用100 nM SignalSilence ® Control siRNA (Unconjugated) #6568， SignalSilence ® p70/85 S6 Kinase siRNA I #6566 (+) 或 SignalSilence ® p70/85 S6 Kinase siRNA II (+)，所用抗体为p70 S6 Kinase (49D7) Rabbit mAb兔单抗 #2708 and α-Tubulin (11H10) Rabbit mAb 兔单抗#2125 。 p70 S6 Kinase (49D7) Rabbit mAb兔单抗验证了p70/85 S6激酶表达的沉默， α-Tubulin (11H10) Rabbit mAb兔单抗作为上样和p70/85 S6 激酶特异性的对照。 |
