| 货号 | 7050C |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit #7050 detects endogenous levels of total p44/42 MAPK protein (see Figure 1). The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 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 | 4C |
| 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. |
Figure 1. Treatment of NIH/3T3 cells with PDGF stimulates phosphorylation of p44/42 MAPK at Thr202/Tyr204, detected by the PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit #7177, but has little effect on the total level of p44/42 MAPK detected by PathScan® Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit. NIH/3T3 cells (80-90% confluent) were starved overnight and treated with 100 ng/mL PDGF for 5 minutes at 37ºC. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots, using p44/42 MAPK (Erk1/2) Antibody #9102 (left panel) and Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (right panel), are shown in the bottom figure.图1：用PDGF 刺激NIH/3T3细胞后使p44/42 MAPK蛋白苏氨酸（202位点）/酪氨酸（204位点）磷酸化，检测试剂盒是PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit #7177，但是Western分析发现总p44/42 MAPK蛋白水平只受到轻微影响，使用的试剂盒是PathScan® Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit。NIH/3T3 cells (80-90% 生长汇合时)无培养基过夜，100 ng/mL PDGF 37ºC 处理5 min。 上图显示450 nm时的OD450读数，下图是对应的Western blot检测结果，使用的抗体是Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370(右侧) 和p44/42 MAPK (Erk1/2) Antibody #9102(左侧)。 | |
Figure 2. The relationship between the protein concentration of the lysate from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm is shown.图2：未处理、PDGF处理的NIH/3T3细胞裂解物中蛋白质浓度和450 nm处光学密度读数之间的线性关系。 |
