| 货号 | 7300C |
| 描述 | CSTs PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-Stat3 (Tyr705) protein. A Stat3 rabbit monoclonal antibody has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-Stat3 proteins are captured by the coated antibody. Following extensive washing, a phospho-Stat3 mouse monoclonal antibody is added to detect the captured phospho-Stat3 protein. HRP-linked anti-mouse 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 phospho-Stat3 protein. |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit detects endogenous levels of Phospho-Stat3 protein. Using this Sandwich ELISA Kit #7300, a significant induction of phospho-Stat3 can be detected in IFN-α-treated HeLa cells. However, the level of total Stat3 protein remains unchanged, as shown by Western analysis (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 | 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 HeLa cells with IFN-alpha stimulates phosphorylation of Stat3 at Tyr705, detected by PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA kit, #7300, but does not affect the level of total Stat3 protein, detected by a Stat3 antibody via Western analysis. OD450 readings are shown in the top figure, while the corresponding Western blot using Phospho-Stat3 (Tyr705) (3E2) Monoclonal Antibody #9138 or Stat3 antibody, is shown in the bottom figure.图1.在HeLa细胞中，用IFN-alpha刺激产生Tyr705位点磷酸化的Stat3,并通过PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA kit#7300检测, 经western免疫印迹检测表明总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Phospho-Stat3 (Tyr705) (3E2) 单克隆抗体#9138 或Stat3 antibody。 | |
Figure 2: Linear relationship between protein concentration of lysates from IFN-alpha-treated HeLa cells and kit assay optical density readings. HeLa cells (75% confluence) were treated with IFN-alpha (100 ng/ml), and lysed after growth at 37oC for 10 min.图2.图示为经IFN-alpha处理的HeLa细胞的裂解液蛋白浓度与试剂盒实验最佳光密度读数的线性关系。HeLa细胞(75% confluence)经过IFN-alpha (100 ng/ml)后在37ºC条件下生长10 min然后裂解。 |
