PathScan? Total β-Catenin Sandwich ELISA Kit

• 产品编号：CST-7308C      品牌：CST       原厂货号：7308C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

CST的 PathScan^® Total Beta-Catenin Sandwich ELISA Kit #7308可以识别内源的总beta-catenin蛋白。如图1所示，本试剂盒可以检测经Calyculin A处理或未处理的细胞裂解液内磷酸化与非磷酸化β-catenin蛋白。如图3 所示，被β-Catenin抗体#9562包被的微孔捕获的蛋白能够在western blot实验中显示出对应β-Catenin蛋白的一条主带。内部测试显示，本试剂盒可以检测来自特定物种的蛋白，但也可能可以识别其他物种的同源蛋白。CST公司的PathScan^® Total Beta-Catenin Sandwich ELISA Kit是基于“三明治”原理的固相酶联免疫吸附试验（ELISA）以检测内源性的总Beta-catenin蛋白。Microwells 已经过Beta-catenin抗体提前包被。使用细胞裂解物孵育后，磷酸化与非磷酸化Beta-catenin蛋白可以被抗体捕获。经过多次冲洗后，Beta-catenin的抗体用于检测被捕获到的Beta-catenin蛋白。辣根过氧化物酶（HRP）标记的抗兔IgG被作为二抗使用。HRP底物（TMB）用于产生颜色。产生的吸光度值与Beta-catenin蛋白含量成正比。β-catenin是Wnt信号通路下游的重要效应分子（1）。在脊椎动物体内它涉及到两个重要的生物过程：早期胚胎发育（2）和肿瘤发生（3）。CK1可以磷酸化β-catenin 45位丝氨酸。该磷酸化将导致β-catenin能够随后被GSK-3磷酸化（4-6）。GSK-3β通过磷酸化β-catenin Ser33, Ser37, 和 Thr41，并促使其进一步被降解（7）。这些位点发生突变会提高β-catenin蛋白稳定性，并且已在多种癌细胞株中发现这些突变（8）。

1. Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305.
2. Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88.
3. Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21.
4. Amit, S. et al. (2002) Genes Dev 16, 1066-76.
5. Liu, C. et al. (2002) Cell 108, 837-47.
6. Yanagawa, S. et al. (2002) EMBO J 21, 1733-42.
7. Yost, C. et al. (1996) Genes Dev 10, 1443-54.
8. Morin, P.J. et al. (1997) Science 275, 1787-90.

原厂资料：

Specificity / Sensitivity

CST''s PathScan^® Total Beta-Catenin Sandwich ELISA Kit #7308 detects endogenous levels of total beta-catenin protein. As shown in Figure 1, both phospho-and nonphospho-β-catenin protein from untreated and Calyculin A-treated 293 cell lysates are detected by this kit. In Figure 3, Western blot analysis of protein captured in the β-Catenin Antibody #9562 coated microwell shows a major band corresponding to the β-catenin 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 Beta-Catenin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Beta-catenin protein. A Beta-Catenin Ab has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-Beta-catenin proteins are captured by the coated antibody. Following extensive washing, Beta-Catenin Mouse mAb is added to detect both the captured phospho- and nonphospho-Beta-catenin protein. Anti-mouse IgG, HRP-linked Antibody #7076 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 Beta-catenin protein. 
 Antibodies in kit are custom formulations specific to kit.

Background

β-Catenin is a key downstream effector in the Wnt signaling pathway (1). It is implicated in two major biological processes in vertebrates: early embryonic development (2) and tumorigenesis (3). CK1 phosphorylates β-catenin at Ser45. This phosphorylation event primes β-catenin for subsequent phosphorylation by GSK-3β (4-6). GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37, and Thr41 (7). Mutations at these sites result in the stabilization of β-catenin protein levels and have been found in many tumor cell lines (8).

1. Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305.
2. Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88.
3. Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21.
4. Amit, S. et al. (2002) Genes Dev 16, 1066-76.
5. Liu, C. et al. (2002) Cell 108, 837-47.
6. Yanagawa, S. et al. (2002) EMBO J 21, 1733-42.
7. Yost, C. et al. (1996) Genes Dev 10, 1443-54.
8. Morin, P.J. et al. (1997) Science 275, 1787-90.

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).