| 货号 | 7976C |
| 描述 | The PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of mTOR protein phosphorylated at Ser2448. A mTOR mouse antibody has been coated onto the microwells. After incubation with cell lysates, mTOR (phospho and nonphospho) protein is captured by the coated antibody. Following extensive washing, a phospho-mTOR (Ser2448) rabbit antibody is added to detect the captured phospho-mTOR 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 mTOR phosphorylated at Ser2448. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse/Rat/Mink |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit #7976 detects endogenous levels of human mTOR protein when phosphorylated at Ser2448 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 mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (1-3) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (4,5). When sufficient nutrients are available, mTOR responds to a phosphatidic acid-mediated signal to transmit a positive signal to p70 S6 kinase and participate in the inactivation of the eIF4E inhibitor, 4E-BP1 (6). These events result in the translation of specific mRNA subpopulations. mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481 (7,8). mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors. For these reasons, mTOR is currently under investigation as a potential target for anti-cancer therapy (9). |
| 存放说明 | 4C |
| 参考文献 | Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22. Brown, E.J. et al. (1994) Nature 369, 756-8. Sabatini, D.M. et al. (1994) Cell 78, 35-43. Gingras, A.C. et al. (2001) Genes Dev 15, 807-26. Dennis, P.B. et al. (2001) Science 294, 1102-5. Fang, Y. et al. (2001) Science 294, 1942-5. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31. Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23. Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7. |
Figure1: Treatment of HeLa cells with Calyculin A accumulates phosphorylation of mTOR at Ser2448, as detected by PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit #7976, but does not affect levels of total mTOR protein detected by PathScan® Total mTOR Sandwich ELISA Kit #7974. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using mTOR (7C10) Rabbit mAb #2983 (left panel) or Phospho-mTOR (Ser2448) Antibody #2971 (right panel) is shown in the bottom figure. 图1：使用Calyculin A 处理HeLa细胞系促使mTOR蛋白在Ser2448位点磷酸化，随后使用PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit #7976检测，但是不影响通过PathScan® Total mTOR Sandwich ELISA Kit #7974检测mTOR总蛋白水平。450 nm吸光度值显示在最上图中。然而通过使用mTOR (7C10)兔源抗体#2983 (左图)和 Phospho-mTOR (Ser2448) Antibody #2971 (右图)进行western blot分析，结果显示在下图中。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and Calyculin A-treated HeLa cells and the absorbance at 450 nm is shown. After starvation, HeLa cells (85% confluence) were treated with Calyculin A (10 nM) for 10-15 min at 37ºC and then lysed. 图2：对照组和Calyculin A 处理组的HeLa细胞裂解物的蛋白浓度与在450 nm吸光度值之间的关系如图所示。饥饿培养后，HeLa细胞(85% confluence)加入Calyculin A (10 nM)，并在37ºC处理10-15分钟，然后裂解。 |
