PathScan® Total β-Catenin Sandwich ELISA Kit【科瑞奥博】

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PathScan® Total β-Catenin Sandwich ELISA Kit

货号： 7308C 基本售价： 6346.0 元规格： 1Kit

产品信息

概述
货号	7308C
描述	CSTs 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.
反应种属	Human/Mouse/Monkey
应用	ELISA
目标/特异性	CSTs 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.

性能
供应商	CST
背景	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).
存放说明	4C
参考文献	Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305. Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88. Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21. Amit, S. et al. (2002) Genes Dev 16, 1066-76. Liu, C. et al. (2002) Cell 108, 837-47. Yanagawa, S. et al. (2002) EMBO J 21, 1733-42. Yost, C. et al. (1996) Genes Dev 10, 1443-54. Morin, P.J. et al. (1997) Science 275, 1787-90.

参考图片

Figure 1: Nonphospho- and phospho-β-catenin proteins from untreated and Calyculin A-treated 293 cells can be detected by PathScan^® Total β-Catenin Sandwich ELISA kit #7308 with similar optical density readings. OD 450 readings are shown in the top figure, while the corresponding Western blot using Phospho-β-Catenin (Ser45) Antibody #9564 (right panel) or β-Catenin Antibody #9562 (left panel), is shown in the bottom figure.

图1：使用PathScan^® Total β-Catenin Sandwich ELISA kit #7308检测未经处理和Calyculin A处理过293细胞的非磷酸化和磷酸化-β-catenin蛋白。OD450读数如图上部所示，相应使用Phospho-β-Catenin (Ser45) 抗体 #9564（右）或β-Catenin 抗体 #9562（左）检测蛋白进行western blot如图下部所示。

Figure 2: The relationship between protein concentration of lysates from untreated and Forskolin treated COS cells and kit assay optical density readings is shown. COS cells were treated with Forskolin #3828 (50 µM) for 30 min at 37°C, and then lysed.

未处理和Forskolin处理的 COS细胞和试剂盒蛋白裂解液的蛋白浓度与OD读数间的关系如图所示。COS细胞经Forskolin #3828 (50 µM) 处理37°C，30 min 后裂解。

Figure 3: Kit specificity demonstrated by Western blot analysis of the ELISA-well captured protein. Lysates were prepared from human 293 cells and incubated in wells coated with capture β-Catenin Antibody #9562. Wells were then washed and captured protein was solubilized in SDS gel loading buffer. Western blot analysis of 293 cell starting lysate (lane 1) and captured protein (lane 2) was performed, using β-Catenin Mouse mAb #2322. The major band detected in the captured material corresponds to the β-catenin protein (lane 2).

图3：试剂盒特异性通过使用western blot分析ELISA捕获到的蛋白以确认。人293细胞裂解物在孔道中与捕获抗体β-Catenin Antibody #9562共同孵育。随后冲洗孔道，捕获的蛋白溶解在SDS胶上样缓冲液。Western blot使用β-Catenin Mouse mAb鼠单抗 #2322分析293细胞起始裂解物（1道和捕获到的蛋白（2道）。捕获组分中主带对应了β-catenin蛋白（2道）。