PathScan? Total Smad2 Sandwich ELISA Kit

• 产品编号：CST-7244C      品牌：CST       原厂货号：7244C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

CST的PathScan^® Total Smad2 Sandwich ELISA Kit #7244可以识别内源性的总Smad2蛋白。如图1所示，本试剂盒可以检测未经TGFβ处理和处理后的MV1LU细胞内磷酸化与非磷酸化Smad2蛋白。图3的结果显示使用Smad2 Rabbit mAb 兔单抗 #3109孵育后微孔捕获的蛋白，可以在western实验中检测到单一对应Smad2蛋白的条带。本试剂盒也可以检测Hela细胞中Smad2蛋白（结果未显示）。经内部测试显示，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的PathScan^® Total Smad2 Sandwich ELISA试剂盒是基于“三明治”原理的固相酶联免疫吸附试验（ELISA）以识别内源性的总Smad2蛋白。微孔已经过Smad2兔多抗#3109*提前包被。与细胞裂解物孵育后，Smad2（磷酸化与非磷酸化）可以被抗体捕获。经过多次冲洗后，Smad2的Mouse mAb鼠单抗#3119*用于检测被捕获到的磷酸化与非磷酸化的Smad2蛋白。辣根过氧化物酶（HRP）标记的抗鼠IgG#7076*被作为二抗使用。HRP底物（TMB）用于显色。产生的吸光度值与总的Smad2含量成正比。 *表示该抗体本试剂盒专用。Smad信号转导分子家族成员是胞内通路中TGF-β信号从细胞表面传送到细胞核的重要组成部分。Smad 可被分为三类：受体调控型Smad（简称R-Smad），主要包括Smad1,2,3,5和8；共同介导型Smad（co-Smad）包括Smad4；还有拮抗型或称抑制型Smad（I-Smads），包括Smad6和Smad7[1-5]。激活的I型受体与特异的R-Smad有关，并能在R-Smad的保守的C端SSXS基序处对其磷酸化。磷酸化的R-Smad与受体分离，再与co-Smad（Smad4）形成异侧复合物，使复合物迁移入核。一旦进入核内，Smad可以各种DNA结合蛋白为目标调控转录反应。[6-8]

1. Heldin, C.H. et al. (1997) Nature 390, 465-71.
2. Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94.
3. Derynck, R. et al. (1998) Cell 95, 737-40.
4. Massagué, J. (1998) Annu Rev Biochem 67, 753-91.
5. Whitman, M. (1998) Genes Dev 12, 2445-62.
6. Wu, G. et al. (2000) Science 287, 92-7.
7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
8. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.

原厂资料：

Specificity / Sensitivity

CST''s 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.

Description

CST''s 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.

Background

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).

1. Heldin, C.H. et al. (1997) Nature 390, 465-71.
2. Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94.
3. Derynck, R. et al. (1998) Cell 95, 737-40.
4. Massagué, J. (1998) Annu Rev Biochem 67, 753-91.
5. Whitman, M. (1998) Genes Dev 12, 2445-62.
6. Wu, G. et al. (2000) Science 287, 92-7.
7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
8. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.

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).