| 货号 | 7789C |
| 描述 | The PathScan® Total Vimentin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total vimentin protein. A Vimentin Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-vimentin protein is captured by the coated antibody. Following extensive washing, a Vimentin Rabbit Detection Antibody is added to detect the captured vimentin protein. 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 optical density for this developed color is proportional to the quantity of vimentin protein. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | The PathScan® Total Vimentin Sandwich ELISA Kit detects endogenous levels of vimentin protein 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 | The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Major types of intermediate filaments are distinguished by their cell-specific expression: cytokeratins (epithelial cells), glial fibrillary acidic protein (GFAP) (glial cells), desmin (skeletal, visceral, and certain vascular smooth muscle cells), vimentin (mesenchyme origin), and neurofilaments (neurons). GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape (1). In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes (2). Research studies have shown that vimentin is present in sarcomas, but not carcinomas, and its expression is examined in conjunction with that of other markers to distinguish between the two (3). Vimentins dynamic structural changes and spatial re-organization in response to extracellular stimuli help to coordinate various signaling pathways (4). Phosphorylation of vimentin at Ser56 in smooth muscle cells regulates the structural arrangement of vimentin filaments in response to serotonin (5,6). Remodeling of vimentin and other intermediate filaments is important during lymphocyte adhesion and migration through the endothelium (7). |
| 存放说明 | 4C |
| 参考文献 | Eng, L.F. et al. (2000) Neurochem Res 25, 1439-51. Goebel, H.H. et al. (1987) Acta Histochem Suppl 34, 81-93. Leader, M. et al. (1987) Histopathology 11, 63-72. Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41. Tang, D.D. et al. (2005) Biochem J 388, 773-83. Fomina, I.G. et al. (1990) Klin Med (Mosk) 68, 125-7. Nieminen, M. et al. (2006) Nat Cell Biol 8, 156-62. Yamaguchi, T. et al. (2005) J Cell Biol 171, 431-6. Oguri, T. et al. (2006) Genes Cells 11, 531-40. Zhu, Q.S. et al. (2011) Oncogene 30, 457-70. Xue, G. and Hemmings, B.A. (2013) J Natl Cancer Inst 105, 393-404. |
Figure 1. Treatment of HeLa cells with paclitaxel stimulates phosphorylation of vimentin at Ser56, while treatment with hydroxyurea reduces that phosphorylation, as detected by the PathScan® Phospho-Vimentin (Ser56) Sandwich ELISA Kit #7795. Neither treatment affects the levels of total vimentin as detected by the PathScan® Total Vimentin Sandwich ELISA Kit #7789. HeLa cells (80-90% confluent) were treated with 100 nM Paclitaxel #9807 or 4 mM hydroxyurea for 20 hr and lysed. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using Vimentin (D21H3) XP® Rabbit mAb #5741 (left panel) or Phospho-Vimentin (Ser56) (D5H2) Rabbit mAb #7391 (right panel) are shown in the bottom figure. 图1，如athScan® Phospho-Vimentin (Ser56) Sandwich ELISA Kit #7795显示，紫杉醇处理HeLa细胞诱导vimentin蛋白56位丝氨酸磷酸化，使用羟基脲处理则会减少磷酸化。任一处理都不会影响vimentin的总蛋白量。使用100nM紫杉醇#9807或4mM羟基脲处理HeLa细胞（80-90%接触）20小时然后裂解细胞。450nm处吸光值如上图所示，使用Vimentin (D21H3) XP®兔 mAb #5741（左）或Phospho-Vimentin (Ser56) (D5H2) Rabbit mAb #7391（右）进行的western blot实验位于下图。 | |
Figure 2. The relationship between the protein concentration of lysates from HeLa cells, treated with Paclitaxel #9807 or hydroxyurea, and the absorbance at 450 nm using the PathScan® Total Vimentin Sandwich ELISA Kit is shown. 图2，使用PathScan® Total Vimentin Sandwich ELISA说明经紫杉醇#9807或羟基脲处理后，HeLa细胞裂解液蛋白浓度与450nm吸光值的关系。 |
