| 货号 | 7946C |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit detects endogenous levels of p38 MAP kinase phosphorylated at Thr180/Tyr182 in human and mouse cells. As shown in Figure 1, a significant induction of p38 MAP kinase phosphorylation at Thr180/Tyr182 can be detected in HeLa cells using the PathScan®Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit following treatment with anisomycin or UV irradiation. The level of total p38 MAP kinase remains unchanged as shown by western analysis (Figure 1). 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 | p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAP kinase, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. Activated p38 MAP kinase has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10). |
| 存放说明 | 4C |
| 参考文献 | Rouse, J. et al. (1994) Cell 78, 1027-37. Han, J. et al. (1994) Science 265, 808-11. Lee, J.C. et al. (1994) Nature 372, 739-46. Freshney, N.W. et al. (1994) Cell 78, 1039-49. Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6. Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4. Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30. Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31. |
Figure 1. Treatment of HeLa cells with anisomycin or UV irradiation stimulates phosphorylation of p38 MAP kinase at Thr180/Tyr182 as detected by the PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit #7946, but does not affect the level of total p38 MAP kinase. HeLa cells were treated with 5 μg/ml anisomycin for 30 minutes at 37ºC or 45 mJ/cm2 UV irradiation followed by a 30 minute recovery period at 37ºC. The absorbance readings at 450 nm are shown in the top figure, while the western blots using p38 MAPK Antibody #9212 (left) or Phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211 (right) are shown in the bottom figure.图1：用茴香霉素或UV辐射刺激HeLa细胞后使p38 MAP kinase蛋白苏氨酸（180位点）/酪氨酸（182位点）磷酸化，检测试剂盒是PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit #7946，但是总p38 MAP kinase蛋白水平并没有受到影响。HeLa细胞用5 μg/ml茴香霉素37ºC处理30 min，或用 45 mJ/cm2 UV 37ºC照射，并恢复30 min。上图显示OD450读数，下图是对应的Western blot检测结果，使用的抗体是 p38 MAPK Antibody #9212 (左侧)、 Phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211(右侧)。 | |
Figure 2. The relationship between lysate protein concentration from untreated, anisomycin-treated or UV-irradiated HeLa cells and the absorbance at 450 nm is shown.图2：未处理、茴香霉素处理或UV照射的HeLa 细胞裂解物中蛋白质浓度和试剂盒分析光学密度读数之间的线性关系。。 |
