| 货号 | 6436S |
| 反应种属 | Human |
| 应用 | TFN |
| 使用方法 | CST recommends transfection with 100 nM p44 MAPK siRNA I 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 |
| 计算分子量 | 44 |
| 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. Castro-Obregón, S. et al. (2004) J Biol Chem 279, 17543-53. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p44 MAPK (Erk1) siRNA I (+), using p44/42 MAPK (Erk1/2) Antibody #9102 and GCK Antibody #3782. The Erk1/2 antibody confirms silencing of Erk1 expression and GCK antibody is used to control for loading and specificity of p44 MAPK (Erk1) siRNA. | |
Western blot分析细胞提取物：转染了100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-)或SignalSilence® p44 MAPK (Erk1) siRNA I (+)的HeLa细胞，使用p44/42 MAPK (Erk1/2)抗体#9102和GCK抗体#3782进行western blot分析。Erk1/2抗体用以确认Erk1表达的沉默，GCK抗体作为内参，控制上样量和p44 MAPK (Erk1) siRNA的特异性。 |
