| 货号 | 7265C |
| 描述 | The PathScan® Total GSK-3β Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of GSK-3β protein. A GSK-3β mouse mAb has been coated onto the microwells. After incubation with cell lysates, GSK-3β (phospho and nonphospho) protein is captured by the coated antibody. Following extensive washing, a GSK-3β rabbit mAb is added to detect the captured GSK-3β 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 the absorbance for this developed color is proportional to the quantity of total GSK-3β. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Total GSK-3β Sandwich ELISA Kit #7265 detects endogenous levels of total human GSK-3β 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 | Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulates glycogen synthesis in response to insulin (1). GSK-3 is a ubiquitously expressed serine/threonine protein kinase that phosphorylates and inactivates glycogen synthase. GSK-3 is a critical downstream element of the PI3K/Akt cell survival pathway whose activity can be inhibited by Akt-mediated phosphorylation at Ser21 of GSK-3α and Ser9 of GSK-3β (2,3). GSK-3 has been implicated in the regulation of cell fate in Dictyostelium and is a component of the Wnt signaling pathway required for Drosophila, Xenopus, and mammalian development (4). GSK-3 has been shown to regulate cyclin D1 proteolysis and subcellular localization (5).糖原合成酶激酶-3（GSK-3）最初作为调节糖原合成酶被认定，响应胰岛素（1）。 |
| 存放说明 | 4C |
| 参考文献 | Welsh, G.I. et al. (1996) Trends Cell Biol 6, 274-9. Srivastava, A.K. and Pandey, S.K. (1998) Mol Cell Biochem 182, 135-41. Cross, D.A. et al. (1995) Nature 378, 785-9. Nusse, R. (1997) Cell 89, 321-3. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511. |
Figure 1. Treatment of NIH/3T3 cells with hPDGF-BB #8912 stimulates phosphorylation of GSK-3β at Ser9, as detected by PathScan® Phospho-GSK-3β (Ser9) Sandwich ELISA Kit #7311, but does not affect levels of total GSK-3β protein detected by PathScan® Total GSK-3β Sandwich ELISA Kit #7265. The absorbance readings at 450 nm are shown in the upper figure, while the corresponding western blots using GSK-3β (27C10) Rabbit mAb #9315 (left panel) or Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb #5558 (right panel) is shown in the lower figure. 图1. 用hPDGF-BB #8912处理NIH/3T3细胞，刺激GSK-3β丝氨酸9位点的磷酸化，用PathScan® Phospho-GSK-3β (Ser9) Sandwich ELISA Kit #7311检测，但不影响用PathScan® Total GSK-3β Sandwich ELISA Kit #7265检测总GSK-3β蛋白。450 nm处吸光度值见上图，对应的western blots实验结果见下图，使用的抗体为GSK-3β (27C10) Rabbit mAb #9315 (左面板)或Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb #5558 (右面板)。 | |
Figure 2. The relationship between protein concentration of lysates from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm is shown. After starvation, NIH/3T3 cells (85% confluence) were treated with hPDGF-BB #8912 (50 ng/ml) for 10-15 minutes at 37ºC and then lysed. 图2. 如图所示，未处理和用DGF处理的NIH/3T3细胞裂解物蛋白浓度与450nm处吸光度值的关系。经饥饿后，NIH/3T3细胞（达85%汇合），用hPDGF-BB #8912 (50 ng/ml) 37ºC处理10-15min，之后进行裂解。 |
