PathScan? Total p21 Waf1/Cip1 Sandwich ELISA Kit

• 产品编号：CST-7167C      品牌：CST       原厂货号：7167C
• 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's PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167能够检测内源性p21 Waf1/Cip1总蛋白。如图1，使用PathScan® Total p21 Waf1/Cip1 Sandwich ELISA Kit #7167检测293, HeLa, K562和 THP1细胞提取物。检测到的p21 Waf1/Cip1总蛋白与western blot方法检测的蛋白水平相对应。如图3，western blot方法检测被捕获的用p21 Waf1/Cip1 antibody预包被的微孔中的蛋白，显示一条p21 Waf1/Cip1蛋白的带。本试剂盒能够检测通过内部测试的特定种属蛋白，但也许能够检测其他种属的同源蛋白。CST's 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。 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)。

1. Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534.
2. Cheng, J. et al. (1999) EMBO J. 18, 1571-1583.
3. Flores-Rozas, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 8655-8659.
4. Wang, Y. and Prives, C. (1995) Nature 376, 88-91.
5. Sheaff, R.J. et al. (2000) Cell 5, 403-410.
6. Pechnick, R.N. et al. (2008) Proc Natl Acad Sci U S A 105, 1358-63.

原厂资料：

Specificity / Sensitivity

CST's 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. 
 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 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. 
 

Background

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).

1. Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534.
2. Cheng, J. et al. (1999) EMBO J. 18, 1571-1583.
3. Flores-Rozas, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 8655-8659.
4. Wang, Y. and Prives, C. (1995) Nature 376, 88-91.
5. Sheaff, R.J. et al. (2000) Cell 5, 403-410.
6. Pechnick, R.N. et al. (2008) Proc Natl Acad Sci U S A 105, 1358-63.

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).