| 货号 | 7995C |
| 描述 | The PathScan® Phospho-Stat3 (Ser727) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat3 protein phosphorylated at Ser727. A Stat3 rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Stat3 (phospho or nonphospho) protein is captured by the coated antibody. Following extensive washing, a phospho-Stat3 (Ser727) mouse mAb is added to detect the captured phospho-Stat3 protein. Anti-mouse 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 Stat3 phosphorylated at Ser727. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-Stat3 (Ser727) Sandwich ELISA Kit detects endogenous levels of Stat3 protein phosphorylated at Ser727 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. |
| 供应商 | CST |
| 背景 | The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8). |
| 存放说明 | 4C |
| 参考文献 | Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85. Bromberg, J.F. et al. (1999) Cell 98, 295-303. Darnell, J.E. et al. (1994) Science 264, 1415-21. Ihle, J.N. (1995) Nature 377, 591-4. Wen, Z. et al. (1995) Cell 82, 241-50. Yokogami, K. et al. (2000) Curr Biol 10, 47-50. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94. |
Figure 1: Treatment of A-431 cells with EGF stimulates phosphorylation of Stat3 at Ser727, as detected by PathScan® Phospho-Stat3 (Ser727) Sandwich ELISA Kit #7995, but does not affect levels of total Stat3 protein detected by PathScan® Total Stat3 Sandwich ELISA Kit #7305. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using Stat3 Antibody #9132 (left panel) or Phospho-Stat3 (Ser727) #9134 (right panel) are shown in the bottom figure. 图1.在A-431细胞中，用EGF刺激产生Ser727位点磷酸化的Stat3,并通过PathScan® Phospho-Stat3 (Ser727) Sandwich ELISA Kit #7995检测。但是经PathScan® Total Stat3 Sandwich ELISA Kit #7305检测表明总Stat3蛋白水平并未受到影响。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Stat3 Antibody #9132(左侧图) 或Phospho-Stat3 (Ser727) #9134(右侧图)。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and EGF-treated A431 cells and the absorbance at 450 nm is shown. After starvation, A431 cells (85% confluence) were treated with EGF (100 ng/ml) for 5 min at 37ºC and then lysed. 图2.图示为未经处理和经EGF处理的A431细胞的裂解液蛋白浓度与450 nm 吸光度的关系。经过饥饿后，A431 细胞(85% confluence)在37ºC条件下经EGF (100 ng/ml)处理5 min 并裂解。 |
