| 货号 | 7251C |
| 描述 | The PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of MARCKS when phosphorylated at Serines 152 and 156. A MARCKS rabbit antibody has been coated onto the microwells. After incubation with cell lysates, MARCKS protein (phosphorylated and nonphospho) is captured by the coated antibody. Following extensive washing, a biotinylated phospho-MARCKS (Ser152/156) rabbit monoclonal detection antibody is added to detect the captured phospho-MARCKS protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of MARCKS phosphorylated at Serines 152 and 156. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit #7251 detects endogenous levels of MARCKS protein only when phosphorylated at Serines 152 and 156 as shown in Figure 1. 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. |
| 供应商 | CST |
| 背景 | Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) is a major PKC substrate expressed in many cell types. MARCKS has been implicated in cell motility, cell adhesion, phagocytosis, membrane traffic, and mitogenesis (1). PKC phosphorylates Ser159, 163, 167, and 170 of MARCKS in response to growth factors and oxidative stress. Phosphorylation at these sites regulates the calcium/calmodulin binding and filamentous (F)-actin cross-linking activities of MARCKS (2-4). Phosphorylation by PKC also results in translocation of MARCKS from the plasma membrane to the cytoplasm (5). |
| 存放说明 | 4C and -20C |
| 参考文献 | Ramsden, J.J. (2000) Int J Biochem Cell Biol 32, 475-9. Heemskerk, F.M. et al. (1993) Biochem Biophys Res Commun 190, 236-41. Graff, J.M. et al. (1989) J Biol Chem 264, 21818-23. Hartwig, J.H. et al. (1992) Nature 356, 618-22. Thelen, M. et al. (1991) Nature 351, 320-2. |
Figure 1. Treatment of Hela cells with TPA #4174 stimulates phosphorylation of MARCKS at Ser152/Ser156, while cells treated with bisindolylmaleimide inhibits phosphorylation, as detected by the PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit #7251. Hela cells (80-90% confluent) were treated with 200 μM TPA for 30 minutes at 37ºC, or treated with 2.0 μM bisindolylmaleimide for 3 hours. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using MARCKS (D88D11) XP® Rabbit mAb #5607 (left panel) or Phospho-MARCKS (Ser152/156) Antibody #2741 (right panel) are shown in the bottom figure. 图1. HeLa细胞经TPA #4174处理，刺激MARCKS Ser152/Ser156的磷酸化，而细胞用双吲哚马来酰亚胺（bisindolylmaleimide）会抑制磷酸化，可用PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit #7251进行检测。用PathScan® Total MARCKS Sandwich ELISA Kit #7188检测，MARCKS总水平不会受两种处理方式的影响。HeLa细胞（80％-90％汇合）用200 μM TPA 37ºC 处理30min，或用2.0 μM 双吲哚马来酰亚胺（bisindolylmaleimide）处理3小时。在450 nm处读取吸光度值，如上图所示。而相应的western blots结果见下图，使用的抗体为MARCKS (D88D11) XP® Rabbit mAb #5607 (左图) 或Phospho-MARCKS (Ser152/156) Antibody #2741 (右图)。 | |
Figure 2. The relationship between the protein concentration of lysates from untreated, bisindolylmaleimide-treated or TPA-treated HeLa cells and the absorbance at 450 nm using the PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit is shown. 图2.如图所示，细胞未处理、用双吲哚马来酰亚胺（bisindolylmaleimide）或TPA处理后细胞裂解物蛋白浓度的关系，并用PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit 检测450nm处的吸光度值。 |
