| 货号 | 7167C |
| 描述 | CSTs PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total p21 Waf1/Cip1 protein. A p21 Waf1/Cip1 mouse mAb * has been coated onto the microwells. After incubation with cell lysates, total p21 Waf1/Cip1 protein is captured by the coated antibody. Following extensive washing, a p21 Waf1/Cip1 antibody* is added to detect the captured total p21 Waf1/Cip1 protein. Anti-rabbit IgG, HRP-linked Antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of total p21 Waf1/Cip1 protein. * Antibodies in kit are custom formulations specific to kit.CSTs PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit为固相三明治酶联免疫吸附试剂盒，能够检测内源性p21 Waf1/Cip1总蛋白。p21 Waf1/Cip1 mouse mAb包被于微孔上。与细胞裂解液孵育后，p21 Waf1/Cip1总蛋白被包被抗体捕获。多次洗涤后，加入p21 Waf1/Cip1 antibody*检测被捕获的p21 Waf1/Cip1总蛋白。随后，用Anti-rabbit IgG, HRP-linked Antibody #7074*识别结合的检测抗体。加入HRP底物(TMB)进行显色。显色过程中吸光度的强度和p21 Waf1/Cip1总蛋白的量成正比。*不同试剂盒中的抗体有不同的组成。 |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167 detects endogenous levels of total p21 Waf1/Cip1 protein. As shown in Figure 1, cell lysates from 293, HeLa, K562 and THP1 are analyzed using PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167. Measured levels of total p21 Waf1/Cip1 protein correlate with levels detected by Western blot analysis. In Figure 3, Western blot analysis of protein captured in the p21 Waf1/Cip1 antibody coated microwell shows a major band corresponding to the p21 Waf1/Cip1 protein. |
| 供应商 | CST |
| 背景 | The tumor suppressor protein p21 Waf1/Cip1 acts as an inhibitor of cell cycle progression. It functions in stoichiometric relationships forming heterotrimeric complexes with cyclins and cyclin-dependent kinases. In association with CDK2 complexes, it serves to inhibit kinase activity and block progression through G1/S (1). However, p21 may also enhance assembly and activity in complexes of CDK4 or CDK6 and cyclin D (2). The carboxy-terminal region of p21 is sufficient to bind and inhibit PCNA, a subunit of DNA polymerase, and may coordinate DNA replication with cell cycle progression (3). Upon UV damage or during cell cycle stages when cdc2/cyclin B or CDK2/cyclin A is active, p53 is phosphorylated and upregulates p21 transcription via a p53-responsive element (4). Protein levels of p21 are downregulated through ubiquitination and proteasomal degradation (5).肿瘤抑制蛋白p21 Waf1/Cip1能够充当细胞周期进程抑制剂。其功能与周期蛋白和周期蛋白依赖性激酶形成的异源三聚体存在化学计量关系。通过与CDK2复合体联合，它可以抑制激酶的活性和阻断G1/S进程(1)。然而，p21也可以增强CDK4或者CDEK6与周期蛋白-D形成的复合体的组装和活性(2)。p21的羧基末端区域能够充分结合并抑制PCNA，DNA聚合酶的一个亚基，而且可以协调细胞周期进程中DNA的复制(3)。基于紫外线损伤或者是在细胞周期中cdc2/cyclin B或CDK2/cyclin A活跃阶段，p53被磷酸化且通过p53-响应元件上调p21的表达(4)。泛素化和蛋白酶体降解能够下调p21的蛋白水平(5)。 |
| 存放说明 | 4C |
| 参考文献 | Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534. Cheng, J. et al. (1999) EMBO J. 18, 1571-1583. Flores-Rozas, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 8655-8659. Wang, Y. and Prives, C. (1995) Nature 376, 88-91. Sheaff, R.J. et al. (2000) Cell 5, 403-410. Pechnick, R.N. et al. (2008) Proc Natl Acad Sci U S A 105, 1358-63. |
Figure 1: p21 Waf1/Cip1 protein from 293, HeLa, K562 and THP1 cells is detected by PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167. The levels of p21 Waf1/Cip1 protein measured using PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167 correlate with p21 Waf1/Cip1 protein levels detected by Western blot analysis. Absorbance450 nm is shown in the top figure, while the corresponding Western blot using p21 Waf1/Cip1 Antibody #2946, is shown in the bottom figure.使用PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167检测293、 HeLa、K562 和THP1 细胞中p21 Waf1/Cip1的蛋白水平。PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167检测的p21 Waf1/Cip1的蛋白水平与western blot方法检测的蛋白水平相对应。上图示450nm处吸光值，对应的western blot使用p21 Waf1/Cip1 Antibody #2946，如下图。 | |
Figure 2: The relationship between protein concentration of lysates from 293 cells and the absorbance450 nm is shown.图2:293细胞裂解液蛋白浓度和450nm吸光度之间的关系。 | |
Figure 3: Kit specificity demonstrated by Western blot analysis of the ELISA-well capture protein. Lysates were prepared from human 293 cells and incubated in wells coated with capture p21 Waf1/Cip1 antibody. Wells were then washed and captured protein was solubilized in SDS gel loading buffer. Western blot analysis of 293 cell starting lysate (lane 1) was performed, using p21 Waf1/Cip1 Antibody #2931. The major band detected in the captured material corresponds to the p21 Waf1/Cip1 protein (lane 2).图3：western blot方法检测ELISA微孔内捕获的蛋白以证明试剂盒的特异性。人293细胞提取物在预包被p21 Waf1/Cip1捕获抗体的微孔内孵育。随后洗涤微孔，将捕获的蛋白溶于SDS胶上样缓冲液中。Western blot方法检测293细胞初始裂解液（泳道1），使用的抗体为p21 Waf1/Cip1 Antibody #2931。微孔中包被蛋白的主带显示为p21 Waf1/Cip1（泳道2）. |
