PathScan? Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit

• 产品编号：CST-12003S      品牌：Cell Signaling Technology       原厂货号：12003S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

描述：

PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit能够识别内源性水平的人细胞总磷酸化Smad3蛋白（丝氨酸（423/425位）），见图1.试剂盒的灵敏性见图2. 本试剂盒用来检测指示物种的蛋白，即通过内部测试确定的那些，也可以用来检测来自其他物种的同源蛋白。PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit是固相夹心酶联免疫吸附试验（ELISA），能够识别内源性水平的磷酸化Smad3蛋白（丝氨酸（423/425位））。微孔包被Smad2/3 鼠抗。细胞裂解液孵育后，Smad3蛋白（磷酸和非磷酸）由包被的抗体捕获。充分冲洗后，加入磷酸-Smad3 (Ser423/425)兔检测抗体检测捕获的磷酸化Smad3蛋白（423/425位）。随后用抗兔IgG, HRP-标记的抗体识别结合的检测蛋白。加入HRP底物TMB进行显色。显色的吸光度值与磷酸化Smad3蛋白（丝氨酸（423/425位））的量成正比。试剂盒中的抗体是针对试剂盒定制的。信号转导分子Smad蛋白家族成员是从细胞表面传输TGF-β信号到细胞核中的一个关键细胞内通路的组成部分。已经定义三种不同类别的Smad蛋白：受体调控Smads（R-Smads），其中包括Smad1，2，3，5和8；共同介质型Smad（Co-Smads），Smad4；拮抗或抑制Smads（I-Smads），Smad6和7（1-5）。激活I型受体与特定的R-Smads联合，并在一个保守的羧基终端SSXS模体将其磷酸化。磷酸化R-Smad从受体解离，并与co-Smad (Smad4)形成杂聚肽复合物，允许复合物转移至细胞核中。一旦达到细胞核内，Smads可以针对各种DNA结合蛋白，调节转录反应（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

PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit recognizes endogenous levels of phospho-Smad3 (Ser423/425) protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that recognizes endogenous levels of Smad3 (Ser423/425) protein. A Smad2/3 Mouse Antibody has been coated on the microwells. After incubation with cell lysates, Smad3 proteins (phospho and nonphospho) are captured by the coated antibody. Following extensive washing, a Phospho-Smad3 (Ser423/425) Rabbit Detection Antibody is added to detect captured phospho-Smad3 (Ser423/425) proteins. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Smad3 (Ser423/425) proteins. 

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