PathScan? Total PRAS40 Sandwich ELISA Kit

• 产品编号：CST-7331C      品牌：CST       原厂货号：7331C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

PathScan ® Total PRAS40 Sandwich ELISA Kit可以检测内源性的总PRAS40蛋白，如图1所示，试剂盒的灵敏度见图2。本试剂盒经内部验证可以检测特定物种的蛋白，但也可能检测其它物种同源性高的蛋白。PathScan ® Total PRAS40 Sandwich ELISA Kit是一种固相三明治酶联免疫吸附方法（ELISA），用来检测内源性的PRAS40 蛋白。兔源PRAS40 抗体已经包被在孔中，与细胞的裂解液共孵育后，PRAS40蛋白被已包被的抗体捕获。经过彻底的洗涤，加入PRAS40 鼠源检测抗体以检测被捕获的PRAS40 蛋白。 HRP标记的抗鼠IgG抗体用来识别被结合的检测抗体。加入HRP底物TMB后显色。颜色的吸光度值与PRAS40 蛋白的含量呈正相关。许多生长因子和激素诱导PI3K激酶信号通路，从而激活下游效应蛋白，例如丝氨酸/苏氨酸激酶Akt（1,2）。一个已知的Akt的底物是一个40 kDa的，富含脯氨酸的蛋白（PRAS40），可以结合14-3-3蛋白（2）。PRAS40还能够结合mTOR，传递Akt信号到mTOR复合物。抑制mTOR信号可以刺激PRAS40结合到mTOR上，反过来抑制mTOR的活性（3）。在缺失胰岛素的细胞内PRAS40和mTOR复合物1（mTORC1）中的Raptor 结合，进而抑制细胞周期蛋白Rheb调节的mTORC1信号通路的激活。Akt磷酸化PRAS40的Thr246位点，解除PRAS40对mTORC1的抑制（4）。反过来mTORC1磷酸化PRAS40的Ser183（5）。

1. Cantley, L.C. (2002) Science 296, 1655-7.
2. Kovacina, K.S. et al. (2003) J Biol Chem 278, 10189-94.
3. Vander Haar, E. et al. (2007) Nat Cell Biol 9, 316-23.
4. Sancak, Y. et al. (2007) Mol Cell 25, 903-15.
5. Oshiro, N. et al. (2007) J Biol Chem 282, 20329-39.

原厂资料：

Specificity / Sensitivity

PathScan® Total PRAS40 Sandwich ELISA Kit #7331 detects endogenous levels of PRAS40 protein as shown in Figure 1. 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 PRAS40 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PRAS40. A PRAS40 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, PRAS40 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a PRAS40 mouse detection antibody is added to detect captured PRAS40 protein. Anti-mouse 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 PRAS40 protein.

Background

Many growth factors and hormones induce the phosphoinositide 3-kinase signaling pathway, which results in the activation of downstream effector proteins such as the serine/threonine kinase Akt (1,2). One known Akt substrate is a 40 kDa, proline-rich protein (PRAS40) that binds to 14-3-3 protein (2). PRAS40 also binds mTOR to transduce Akt signals to the mTOR complex. Inhibition of mTOR signaling stimulates PRAS40 binding to mTOR, which in turn inhibits mTOR activity (3). PRAS40 interacts with Raptor in mTOR complex 1 (mTORC1) in insulin-deprived cells and inhibits the activation of the mTORC1 pathway mediated by the cell cycle protein Rheb. Phosphorylation of PRAS40 by Akt at Thr246 relieves PRAS40 inhibition of mTORC1 (4). mTORC1 in turn phosphorylates PRAS40 at Ser183 (5).

1. Cantley, L.C. (2002) Science 296, 1655-7.
2. Kovacina, K.S. et al. (2003) J Biol Chem 278, 10189-94.
3. Vander Haar, E. et al. (2007) Nat Cell Biol 9, 316-23.
4. Sancak, Y. et al. (2007) Mol Cell 25, 903-15.
5. Oshiro, N. et al. (2007) J Biol Chem 282, 20329-39.

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).