| 货号 | 7325C |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Kit detects endogenous levels of Phospho-SAPK/JNK protein. Using this Sandwich ELISA Kit #7325, a significant induction of phospho-SAPK/JNK in 293 cells treated with UV is detected. However, the level of total SAPK/JNK (phospho and non-phospho), detected by PathScan® Total SAPK/JNK Sandwich ELISA Kit #7330, remains unchanged (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 | The stress-activated protein kinase/Jun-amino-terminal kinase SAPK/JNK is potently and preferentially activated by a variety of environmental stresses including UV and gamma radiation, ceramides, inflammatory cytokines, and in some instances, by growth factors and GPCR agonists (1-6). As with the other MAPKs, the core signaling unit is composed of a MAPKKK, typically MEKK1-MEKK4, or by one of the mixed lineage kinases (MLKs), which phosphorylate and activate MKK4/7. Upon activation, MKKs phosphorylate and activate the SAPK/JNK kinase (2). Stress signals are delivered to this cascade by small GTPases of the Rho family (Rac, Rho, cdc42) (3). Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs (3). Alternatively, MKK4/7 can be activated in a GTPase-independent mechanism via stimulation of a germinal center kinase (GCK) family member (4). There are three SAPK/JNK genes each of which undergoes alternative splicing resulting in numerous isoforms (3). SAPK/JNK, when active as a dimer, can translocate to the nucleus and regulate transcription through its effects on c-Jun, ATF-2, and other transcription factors (3,5). |
| 存放说明 | 4C |
| 参考文献 | Davis, R.J. (1999) Biochem Soc Symp 64, 1-12. Ichijo, H. (1999) Oncogene 18, 6087-93. Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69. Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5. |
Figure 1: Treatment of 293 cells with UV stimulates phosphorylation of SAPK/JNK at Thr202/Tyr204, detected by PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA kit #7325, but does not affect the level of total SAPK/JNK protein detected by PathScan® Total SAPK/JNK Sandwich ELISA kit #7330. OD450 readings are shown in the top figure, while the corresponding western blot using Phospho-SAPK/JNK (Thr183/Tyr185) Antibody #9251 (right panel) or SAPK/JNK Rabbit mAb #9258 (left panel), is shown in the bottom figure.用紫外照射293细胞后使SAPK/JNK 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-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA kit #7325，但是总SAPK/JNK 蛋白水平并没有受到影响，检测试剂盒是PathScan® Total SAPK/JNK Sandwich ELISA kit #7330。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Phospho-SAPK/JNK (Thr183/Tyr185) Antibody #9251 (右侧) 和SAPK/JNK Rabbit mAb #9258 (左侧)。 | |
Figure 2: Linear relationship between protein concentration of lysates from control and UV-treated 293 cells and kit assay optical density readings. 293 cells (80% confluence) were treated with UV and lysed after incubation at 37oC for 30 minutes.图2：对照组和紫外照射的293细胞裂解物中蛋白质浓度之间的线性关系和试剂盒分析光学密度读数。紫外照射的293细胞（80%生长汇合）37ºC孵育30min后再裂解。 |
