| 货号 | 7145C |
| 反应种属 | Human/Mouse/Rat |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II detects endogenous levels of Phospho-c-Jun (Ser63) protein. As shown in Figure 1, using this Sandwich ELISA Kit #7145, a significant induction of phospho-c-Jun (Ser63) in UV-treated 293 cells can be detected. However, the level of total c-Jun protein (phospho and non-phospho), detected by the Total c-Jun Sandwich ELISA Kit II #7150, remains unchanged. Both C6 and NIH/3T3 cells treated with UV light or anisomycin show similar results (data not shown). |
| 供应商 | CST |
| 背景 | c-Jun is a member of the Jun Family containing c-Jun, JunB and JunD, and is a component of the transcription factor AP-1 (activator protein-1). AP-1 is composed of dimers of Fos, Jun and ATF family members and binds to and activates transcription at TRE/AP-1 elements (reviewed in 1). Extracellular signals including growth factors, chemokines and stress activate AP-1-dependent transcription. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 through SAPK/JNK (reviewed in 2). Knock-out studies in mice have shown that c-Jun is essential for embryogenesis (3), and subsequent studies have demonstrated roles for c-Jun in various tissues and developmental processes including axon regeneration (4), liver regeneration (5) and T cell development (6). AP-1 regulated genes exert diverse biological functions including cell proliferation, differentiation, and apoptosis, as well as transformation, invasion and metastasis, depending on cell type and context (7-9). Other target genes regulate survival as well as hypoxia and angiogenesis (8,10). c-Jun has emerged as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, as well as rheumatoid arthritis (11,12). |
| 存放说明 | 4C |
| 参考文献 | Jochum, W. et al. (2001) Oncogene 20, 2401-12. Davis, R.J. (2000) Cell 103, 239-52. Hilberg, F. et al. (1993) Nature 365, 179-81. Raivich, G. et al. (2004) Neuron 43, 57-67. Behrens, A. et al. (2002) EMBO J 21, 1782-90. Riera-Sans, L. and Behrens, A. (2007) J Immunol 178, 5690-700. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6. Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3. Karamouzis, M.V. et al. (2007) Mol Cancer Res 5, 109-20. Kim, S. and Iwao, H. (2003) J Pharmacol Sci 91, 177-81. Dass, C.R. and Choong, P.F. (2008) Pharmazie 63, 411-4. |
Figure 1: Treatment of 293 cells with UV stimulates phosphorylation of c-Jun at Ser63, detected by PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II #7145, but does not affect the level of total c-Jun protein detected by PathScan® Total c-Jun Sandwich ELISA Kit II #7150. Lambda protein phosphatase (LPP) treatment of control cell lysates (37ºC for 90 minutes) abolished the basal phosphorylation of c-Jun in control lysates shown in both Sandwich ELISA and Western analysis. The OD450 readings are shown in the top figure, while the corresponding Western blot, using Phospho-c-Jun (Ser63) Rabbit mAb #2378 (right panel) or c-Jun Rabbit mAb (6H5) #2374 (left panel), is shown in the bottom figure.图1：用紫外照射293细胞后使c-Jun蛋白丝氨酸（63位点）磷酸化，检测试剂盒是PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II #7145，但是总c-Jun蛋白水平并没有受到影响，检测试剂盒是PathScan® Total c-Jun Sandwich ELISA Kit II #7150。Sandwich ELISA 和Western检测分析都显示用Lambda蛋白磷酸酶（LPP）处理对照细胞裂解物（37ºC，90 min）后，消除了对照裂解物中c-Jun蛋白的基础磷酸化。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-c-Jun (Ser63) Rabbit mAb #2378 (右侧) 和 c-Jun Rabbit mAb (6H5) #2374 (左侧)。 | |
Figure 2: Linear relationship between protein concentration of lysates from UV-treated 293 cells and kit assay optical density readings. Pathscan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II #7145 is more sensitive than kit # 7260 over a range of lysate concentrations. 293 cells (70-90% confluence) were treated without or with UV and lysed after incubation at 37oC for 30 minutes.图2：紫外照射的293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。对于一系列裂解物浓度检测，Pathscan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II #7145比kit # 7260更灵敏性。未经处理和紫外照射的293细胞（70-90%生长汇合）37ºC孵育30min后再裂解。 |
