| 货号 | 7234C |
| 描述 | The PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat1 when phosphorylated at Tyr701. A Stat1 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, Stat1 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, biotinylated Phospho-Stat1 (Tyr701) Rabbit Detection Antibody is added to detect phosphorylation of Tyr701 on the captured Stat1 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 this developed color is proportional to the quantity of Stat1 phosphorylated at Tyr701. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit detects endogenous levels of Phospho-Stat1 when phosphorylated at Tyr701. As shown in Figure 1, a significant induction of Stat1 phosphorylation at Tyr701 can be detected in HeLa cells following treatment with Interferon-γ (IFN-γ) using the Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234. The level of total Stat1 (phospho and nonphospho) remains unchanged as shown by Western analysis. 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 Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation. |
| 存放说明 | 4C |
| 参考文献 | Heim, M.H. (1999) J Recept Signal Transduct Res 19, 75-120. Durbin, J.E. et al. (1996) Cell 84, 443-50. Meraz, M.A. et al. (1996) Cell 84, 431-42. Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7. Frank, D.A. (1999) Mol Med 5, 432-56. Wen, Z. et al. (1995) Cell 82, 241-50. |
Figure 1. Treatment of HeLa cells with IFN-γ stimulates phosphorylation of Stat1 at Tyr701, detected by the PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234, but does not affect the level of total Stat1 protein detected by Western analysis. HeLa cells (80-90% confluent) were starved for 6 hours and treated with 100 ng/mL IFN-γ for 20 minutes at 37oC. The absorbance readings at 450 nm are shown in the top figure, while the corresponding Western blots, using Stat1 (42H3) Rabbit mAb (Human Specific) #9175 (left panel) or Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 (right panel), are shown in the bottom figure.图1.PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234检测经IFN-γ处理诱导Tyr701位点磷酸化Stat1蛋白，但是通过Western免疫印迹实验表明总Stat1蛋白并未受到影响。HeLa细胞(80-90% confluent) 经过6小时的饥饿，在37oC条件下，用 100 ng/mL IFN-γ处理20min。OD450的读数在图片的最上面显示,同时对应的Western免疫印迹在下面图片显示，所用抗体为Stat1 (42H3) Rabbit mAb (Human Specific) #9175(左侧图) 或Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167(右侧图)。 | |
Figure 2. The relationship between lysate protein concentration from untreated and IFN-γ -treated HeLa cells and the absorbance at 450 nm is shown.图2..图示为未经处理和经IFN-γ 处理的HeLa细胞的裂解液蛋白浓度与450 nm 吸光度的关系。 |
