PathScan® Total Smad2 Sandwich ELISA Kit【科瑞奥博】

产品中心

当前位置：首页>产品中心

PathScan® Total Smad2 Sandwich ELISA Kit

货号： 7244C 基本售价： 6346.0 元规格： 1Kit

产品信息

概述
货号	7244C
描述	CSTs PathScan^® Total Smad2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total Smad2 protein. A Smad2 Rabbit mAb #3109* has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-Smad2 proteins are captured by the coated antibody. Following extensive washing, Smad2 Mouse mAb #3119* is added to detect both the captured phospho- and nonphospho-Smad2 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 Smad2 protein.   * Antibodies in kit are custom formulations specific to kit.
反应种属	Human/Mouse/Mink
应用	ELISA
目标/特异性	CSTs PathScan® Total Smad2 Sandwich ELISA Kit #7244 detects endogenous levels of total Smad2 protein. As shown in Figure 1, both phospho- and nonphospho-Smad2 proteins from untreated and TGFβ treated MV1LU cell lysates are detected by this kit. In Figure 3, Western blot analysis of protein captured in the Smad2 Rabbit mAb #3109 coated microwell shows a single band corresponding to the Smad2 protein. Smad2 in Hela cells also can be detected by this kit (data not shown). 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	Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the receptor-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, and 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved carboxy terminal SSXS motif. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6-8).
存放说明	4C
参考文献	Heldin, C.H. et al. (1997) Nature 390, 465-71. Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94. Derynck, R. et al. (1998) Cell 95, 737-40. Massagué, J. (1998) Annu Rev Biochem 67, 753-91. Whitman, M. (1998) Genes Dev 12, 2445-62. Wu, G. et al. (2000) Science 287, 92-7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.

参考图片

Figure 1: Nonphospho and phospho-Smad2 proteins from untreated and TGF-β treated Mv1Lu cells detected by PathScan^® Total Smad2 Sandwich ELISA kit #7244 showing similar optical density readings. OD 450 readings are shown in the top figure, while the corresponding Western blots using Smad2 Mouse mAb #3103 (left panel) or Phospho-Smad2 (Ser465/467) Antibody #3101 (right panel), are shown in the bottom figure.

图1：使用PathScan^® Total Smad2 Sandwich ELISA kit #7244检测未处理和经过TGF-β处理的Mv1Lu细胞中非磷酸化和磷酸化-Smad2蛋白进行检测。450nm处吸光值如图上部所示，相应使用Smad2鼠单抗#3103（左）或磷酸化-Smad2 (Ser465/467)抗体#3101（右）检测蛋白进行western blot如下图所示。

Figure 2: The relationship between protein concentration of lysates from untreated and TGF-β-treated Mv1Lu cells and kit assay optical density readings. After starvation, Mv1Lu cells (85% confluence) were treated with TGF-β (100 ng/ml) for 15-30 min at 37°C, and then lysed.

图2：未处理和经过 TGF-β处理后Mv1Lu细胞提取物中蛋白浓度与试剂盒测得吸光值间关系。饥饿处理后的Mv1Lu 细胞（85%接触抑制）使用37°C处理15-30分钟，然后裂解。

Figure 3: Kit specificity demonstrated by Western blot analysis of the ELISA well captured protein is shown. Lysates were prepared from mink Mv1Lu cells and incubated in wells coated with capture antibody #3109. Wells were then washed, and captured protein was solubilized in SDS gel loading buffer. Mv1Lu lysate (lane 1) and captured protein (lane 2) were analyzed by Western blot using Smad2 Mouse mAb #3119. A single band corresponding to the Smad2 protein is detected in the captured material (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。mink Mv1Lu细胞裂解物在孔道中与捕获抗体#3109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。用Smad2 Mouse mAb 鼠单抗#3119对Mv1Lu细胞裂解物（1道）和捕获到的蛋白（2道）进行western blot分析。