PathScan? Total c-Jun Sandwich ELISA Kit II

• 产品编号：CST-7150C      品牌：CST       原厂货号：7150C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

CST的 PathScan® Total c-Jun Sandwich ELISA Kit II 能检测内源性c-Jun总蛋白的水平。如图1所示，使用 Sandwich ELISA Kit #7145，能够检测到经紫外照射的293细胞中phospho-c-Jun (Ser63)的显著增加。同时，用Total c-Jun Sandwich ELISA Kit II #7150检测c-Jun总蛋白水平（磷酸化与非磷酸化的），保持不变。当检测经紫外照射和茴香霉素处理的C6、 NIH/3T3细胞时也得到了类似的结果（数据未列出）。该试剂盒比#7270试剂盒更灵敏（如图2）。经内部测试确认，本试剂盒可以检测到所列物种中的蛋白。但对其他物种中的同源蛋白可能也可以检测。CST的PathScan® Total c-Jun Sandwich ELISA Kit II是通过固相夹心酶联免疫吸附法（ELISA）来检测内源性的 c-Jun蛋白水平。先将c-Jun rabbit mAb (#2374*)兔单抗包被在微孔板上。与细胞裂解物孵育后，c-Jun蛋白便可被包被的抗体捕获。清洗彻底后，加入c-Jun Mouse mAb 鼠单抗检测c-Jun蛋白水平。再加入Anti-mouse IgG, HRP偶联抗体作为二抗来识别之前绑定的检测抗体。最后加入HRP底物—— TMB，进行显色。产生的吸光值与c-Jun蛋白的数量成正比。*试剂盒中抗体是专为本试剂盒特制的。c-Jun是Jun家族的一个成员，Jun家族成员包括c-Jun, JunB, 和 JunD。c-Jun是转录因子激活蛋白-1（AP-1）的一个组分。AP-1是由Fos和Jun形成的二聚体以及 ATF家族成员组成的，能够结合并激活TRE/AP-1元件的转录（1）。包括生长因子，细胞因子和压力在内的细胞外信号能够激活AP-1依赖的转录过程。C-Jun的转录活性是通过SAPK/JNK对其中的Ser63和Ser73的磷酸化来调控的（2）。小鼠敲除实验表明c-Jun对于胚胎形成是至关重要的（3），进一步的研究发现c-Jun在多种组织和发育过程中发挥作用，包括轴突再生（4），肝脏再生（5）和T细胞发育（6）。AP-1所调控的基因发挥了多种生物学功能，包括细胞增殖、分化、凋亡、转化，侵润和转移，这依赖于细胞种类和具体情况（7-9）。其他的AP-1所调控的靶基因还能够调节细胞存亡，以及低氧和血管生成（8,10）。研究提示c-Jun可以作为癌症、血管重构、急性炎症反应和风湿性关节炎的重要治疗靶点（11,12）。

1. Jochum, W. et al. (2001) Oncogene 20, 2401-12.
2. Davis, R.J. (2000) Cell 103, 239-52.
3. Hilberg, F. et al. (1993) Nature 365, 179-81.
4. Raivich, G. et al. (2004) Neuron 43, 57-67.
5. Behrens, A. et al. (2002) EMBO J 21, 1782-90.
6. Riera-Sans, L. and Behrens, A. (2007) J Immunol 178, 5690-700.
7. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
8. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6.
9. Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3.
10. Karamouzis, M.V. et al. (2007) Mol Cancer Res 5, 109-20.
11. Kim, S. and Iwao, H. (2003) J Pharmacol Sci 91, 177-81.
12. Dass, C.R. and Choong, P.F. (2008) Pharmazie 63, 411-4.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Total c-Jun Sandwich ELISA Kit II detects endogenous levels of total c-Jun protein. As shown in Figure 1, using PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II #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 this Sandwich ELISA Kit #7150, remains unchanged. Both C6 and NIH/3T3 cells treated with UV light or anisomycin show similar results (data not shown). 
 This kit is more sensitive than kit #7270 (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.

Description

CST's PathScan® Total c-Jun Sandwich ELISA Kit II is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total c-Jun protein. A c-Jun rabbit mAb (#2374*) has been coated onto the microwells. After incubation with cell lysates, c-Jun protein is captured by the coated antibody. Following extensive washing, c-Jun Mouse mAb is added to detect the captured c-Jun protein. HRP-linked anti-mouse antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total c-Jun protein. 
 * Antibodies in this kit are custom formulations specific to the kit.

Background

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).

1. Jochum, W. et al. (2001) Oncogene 20, 2401-12.
2. Davis, R.J. (2000) Cell 103, 239-52.
3. Hilberg, F. et al. (1993) Nature 365, 179-81.
4. Raivich, G. et al. (2004) Neuron 43, 57-67.
5. Behrens, A. et al. (2002) EMBO J 21, 1782-90.
6. Riera-Sans, L. and Behrens, A. (2007) J Immunol 178, 5690-700.
7. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
8. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6.
9. Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3.
10. Karamouzis, M.V. et al. (2007) Mol Cancer Res 5, 109-20.
11. Kim, S. and Iwao, H. (2003) J Pharmacol Sci 91, 177-81.
12. Dass, C.R. and Choong, P.F. (2008) Pharmazie 63, 411-4.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).