| 货号 | 7348C |
| 描述 | The PathScan® Phospho-Smad2 (Ser465/467) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Smad2 when phosphorylated at Ser465/467. A Smad2 Mouse Antibody has been coated onto the microwells. After incubation with cell lysates, Smad2 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-Smad2 (Ser465/467) Detection Antibody is added to detect serine phosphorylation of the captured Smad2 protein. Anti-rabbit IgG, HRP-Linked Ab 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 Smad2 phosphorylated at Ser465/467. 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® Phospho-Smad2 (Ser465/467) Sandwich ELISA Kit #7348 detects Smad2 when phosphorylated at Ser465/467. As shown in Figure 1, a significant induction of Smad2 phosphorylation at Ser465/467 can be detected in TGF-β-treated HaCaT cells using the PathScan® Phospho-Smad2 (Ser465/467) Sandwich ELISA Kit #7348. The level of total Smad2 (phospho and nonphospho) remains unchanged as shown by western analysis and by PathScan® Total Smad2 Sandwich ELISA Kit #7244 (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 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. |
Figure 1. Treatment of HaCaT cells with TGF-β stimulates phosphorylation of Smad2 at Ser465/467, detected by PathScan® Phospho-Smad2 (Ser465/467) Sandwich ELISA Kit #7348, but does not affect the level of total Smad2 protein detected by PathScan® Total Smad2 Sandwich ELISA Kit #7244. The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blots using Smad2 (86F7) Rabbit mAb #3122 (left panel) or Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (right panel) are shown in the bottom figure. 图1：TGF-β处理HaCaT细胞诱导Smad2在Ser465/467的磷酸化可以用PathScan® Phospho-Smad2 (Ser465/467) Sandwich ELISA Kit #7348检测，试剂盒检测显示总Smad2含量不变。450 nm处吸光值如图上不所示，对应地使用Smad2 (86F7)Rabbit mAb 兔单抗#3122（左道）或Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb 兔单抗#3108（右道）进行western blot实验的结果如图下部所示。 | |
Figure 2. The relationship between the protein concentration of lysates from untreated and TGF-β-treated HaCaT cells and the absorbance at 450 nm is shown. Cells (85% confluence) were treated with 10 ng/ml TGF-β for 30 min. at 37oC. 未处理和经过TGF-β处理HaCaT细胞的蛋白浓度与450nm吸光值间关系如图所示。细胞（85%接触抑制）在37°C使用10 ng/ml TGF-β处理30分钟. |
