PathScan? Phospho-mTOR (Ser2448) Sandwich ELISA Kit

• 产品编号：CST-7976C      品牌：CST       原厂货号：7976C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit #7976仅检测在Ser2448位点磷酸化的内源性人源mTOR蛋白（如图1所示）。该试剂盒的灵敏度如图2所示。通过内部实验，该试剂盒可检测指定的物种，但是可能也会检测其它物种的同源性蛋白。该PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit是一个固相夹心酶联免疫吸附法（ELISA），其能检测在Ser2448位点磷酸化的内源性mTOR蛋白水平。一个mTOR 鼠源抗体已经被包被在微孔板上。在与细胞裂解液一起孵育后，mTOR (磷酸和非磷酸的)蛋白被包被的抗体吸附。在充分清洗后，一个phospho-mTOR (Ser2448)兔源抗体被加入以检测已被吸附的磷酸化mTOR蛋白。然后，Anti-rabbit IgG, HRP连接的抗体被用于识别已结合的检测抗体。加入HRP底物TMB显色。该颜色的吸光度值的高低与在Ser2448位点磷酸化的mTOR蛋白量多少成正比。试剂盒里的抗体是专门针对该试剂盒定制的。Mammalian target of rapamycin (mTOR, FRAP, RAFT)是一个丝氨酸/苏氨酸蛋白激酶(1-3)，该激酶的功能作为一个ATP和氨基酸的感受器以平衡营养物的供给情况和细胞生长(4,5)。当足够的营养充足时，mTOR响应一个磷脂酸介导的信号并传送一个正向的信号到p70 S6激酶，并且参与eIF4E抑制因子4E-BP1的失活(6)。这些事件导致特异性mRNA亚群的翻译。mTOR蛋白通过PI3 kinase/Akt信号通路在Ser2448位点磷酸化，并可在Ser2481位点发生自磷酸化(7,8)。mTOR蛋白在细胞生长和体内平衡中起到重要作用，并且在肿瘤中受到异常调节。基于这些原因，mTOR蛋白目前作为抗癌治疗靶点正被深入研究(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：

Specificity / Sensitivity

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.

Description

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.

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).