| 货号 | 6211S |
| 描述 | SignalSilence® Akt siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Akt 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 | SignalSilence® Akt siRNA I will inhibit human, mouse and rat Akt1 and Akt2 expression. It will not affect Akt3 expression. |
| 供应商 | CST |
| 背景 | Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip (15) and p21 Waf1/CIP1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).Small interfering RNA (siRNA) has been used to specifically silence Akt in CHO cells and 3T3-L1 adipocytes (20). |
| 存放说明 | -20C |
| 计算分子量 | 60 |
| 参考文献 | Franke, T.F. et al. (1997) Cell 88, 435-7. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602. Franke, T.F. et al. (1995) Cell 81, 727-36. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51. Sarbassov, D.D. et al. (2005) Science 307, 1098-101. Jacinto, E. et al. (2006) Cell 127, 125-37. Cardone, M.H. et al. (1998) Science 282, 1318-21. Brunet, A. et al. (1999) Cell 96, 857-68. Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5. Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8. Hajduch, E. et al. (2001) FEBS Lett 492, 199-203. Cross, D.A. et al. (1995) Nature 378, 785-9. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511. Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30. Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31. Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57. Manning, B.D. et al. (2002) Mol Cell 10, 151-62. Katome, T. et al. (2003) J Biol Chem 278, 28312-23. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Akt siRNA I (+), using Akt Antibody #9272 and p42 MAP Kinase (Erk2) Antibody #9108. Akt antibody confirms silencing of Akt expression, while the p42 MAP kinase (Erk2) antibody is used to control for loading and specificity of Akt siRNA. Western blot 分析分别转染了 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) 或 SignalSilence® Akt siRNA I (+)的Hela细胞的提取物，所用抗体为Akt Antibody 多抗#9272 和 p42 MAP Kinase (Erk2) Antibody #9108.。Akt抗体确定了Akt表达的沉默， p42 MAP kinase (Erk2) antibody作为Akt siRNA的对照。 | |
Western blot analysis of extracts from CHO cells, transfected with non-targeted (-) or SignalSilence® Akt siRNA I (+) siRNA, using Akt Antibody #9272 and p42 MAP Kinase (Erk2) Antibody #9108. The Akt antibody confirms silencing of protein expression while the p42 MAP Kinase (Erk2) antibody was used to control for loading and specificity of Akt siRNA (A). Phospho-GSK-3β (Ser9) Antibody #9336 was used to confirm downstream pathway inhibition (B). Western blot分析分别转染了non-targeted (-) or SignalSilence ® Akt siRNA I (+) siRNA的CHO细胞的提取物，所用抗体为Akt Antibody #9272 和p42 MAP Kinase (Erk2) Antibody #9108。Akt抗体确定了Akt表达的沉默， p42 MAP kinase (Erk2) antibody作为Akt siRNA的对照。Phospho-GSK-3β (Ser9) Antibody #9336用于确定下游信号的抑制（B）。 |
