| 货号 | 6560S |
| 反应种属 | Human |
| 应用 | TFN |
| 使用方法 | CST recommends transfection with 100 nM p44/42 (Erk1/Erk2) siRNA 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. |
| 供应商 | CST |
| 背景 | Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3) and is an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family as well as Mos and Tpl2/Cot. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors such as U0126 and PD98059. |
| 存放说明 | -20C |
| 参考文献 | Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44. Baccarini, M. (2005) FEBS Lett 579, 3271-7. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505. Marais, R. et al. (1993) Cell 73, 381-93. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13. |
Western blot analysis of extracts from Hek 293 cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p44/42 MAPK (Erk1/2) siRNA (+), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 and α-Tubulin (11H10) Rabbit mAb #2125. The p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb confirms silencing of p44/42 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p44/42 MAPK (Erk1/2) siRNA.Western blot方法检测细胞提取物：转染100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) 和SignalSilence® p44/42 MAPK (Erk1/2) siRNA (+)的HeLa细胞。使用的抗体是p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 和α-Tubulin (11H10) Rabbit mAb #2125。p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb证实了p44/42表达沉默，α-Tubulin (11H10) Rabbit mAb被当作负载及p44/42 MAPK (Erk1/2) siRNA的特异性控制来使用。 |
