PathScan? Total mTOR Sandwich ELISA Kit

• 产品编号：CST-7974C      品牌：CST       原厂货号：7974C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

PathScan ® Total mTOR Sandwich ELISA Kit #7974可以检测内源性ｍTOR总蛋白，如图1所示，试剂盒的灵敏度见图2。本试剂盒经过内部验证可以检测特定物种的蛋白， 但可能检测其它物种同源性高的蛋白。PathScan ® Total mTOR (Ser2481) Sandwich ELISA Kit是一种固相三明治酶联免疫吸附方法（ELISA），用来检测在Ser2481位点磷酸化的内源性的mTOR蛋白。鼠源mTOR抗体已经包被在孔中，与细胞的裂解液共孵育后，mTOR蛋白被已包被的抗体捕获。经过彻底的洗涤，加入Phospho-mTOR (Ser2481) Rabbit mAb捕获mTOR蛋白。HRP标记的抗兔IgG抗体用来识别结合的抗体。HRP底物TMB加入后显色。颜色的吸光度值与Ser2481位点磷酸化的mTOR蛋白的含量呈正相关。哺乳动物雷帕霉素（rapamycin）靶蛋白（mTOR，FRAP，RAFT）是一种丝氨酸/苏氨酸蛋白激酶（1-3），其作为ATP和氨基酸的传感器，可以有效调控营养供应及细胞的生长（4,5）。当营养充分时，mTOR响应磷脂酸介导的信号，进而正向调节p70 S6 激酶，并参与失活eIF4E的抑制剂4E-BP1。这些信号导致特定的mRNA亚群的翻译。mTOR可以被Akt信号通路的PI3K激酶磷酸化修饰Ser2448位点，也能自我磷酸化Ser2481位点（7,8）。mTOR的在细胞生长和动态平衡中起着关键作用，在肿瘤中可能存在调控失常。因此，目前很多研究将mTOR作为一个潜在的抗肿瘤治疗靶点（9）。

1. Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
2. Brown, E.J. et al. (1994) Nature 369, 756-8.
3. Sabatini, D.M. et al. (1994) Cell 78, 35-43.
4. Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
5. Dennis, P.B. et al. (2001) Science 294, 1102-5.
6. Fang, Y. et al. (2001) Science 294, 1942-5.
7. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
8. Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
9. Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.

原厂资料：

pecificity / Sensitivity

PathScan® Total mTOR Sandwich ELISA Kit #7974 detects endogenous levels of total human mTOR 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.

Description

The PathScan® Total mTOR Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of mTOR protein. A mTOR mouse antibody has been coated onto the microwells. After incubation with cell lysates, mTOR protein is captured by the coated antibody. Following extensive washing, an mTOR rabbit antibody is added to detect captured 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 protein.

Background

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).

1. Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
2. Brown, E.J. et al. (1994) Nature 369, 756-8.
3. Sabatini, D.M. et al. (1994) Cell 78, 35-43.
4. Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
5. Dennis, P.B. et al. (2001) Science 294, 1102-5.
6. Fang, Y. et al. (2001) Science 294, 1942-5.
7. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
8. Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
9. Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).