| 货号 | 7285C |
| 描述 | CSTs PathScan® Phospho-PTEN (Ser380) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-PTEN (Ser380) protein. A PTEN Mouse mAb* has been coated onto the microwells. After incubation with cell lysates, PTEN protein (phospho and non-phospho) is captured by the coated antibody. Following extensive washing, Phospho-PTEN (Ser380) Antibody* is added to detect the captured phospho-PTEN (Ser380) protein. Anti-rabbit IgG, HRP-linked Antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of phospho-PTEN (Ser380) protein. * Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human/Mouse/Monkey |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-PTEN (Ser380) Sandwich ELISA Kit detects endogenous levels of phospho-PTEN (Ser380) protein. As shown in Figure 1 using the Phospho-PTEN (Ser380) Sandwich ELISA Kit #7285 a high level of phospho-PTEN (Ser380) is detected in HeLa cells. These levels are significantly reduced in HeLa cell lysates treated with λ phosphatase. 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 | PTEN (phosphatase and tensin homologue deleted on chromosome ten), also referred to as MMAC (mutated in multiple advanced cancers) phosphatase, is a tumor suppressor implicated in a wide variety of human cancers (1). PTEN encodes a 403 amino acid polypeptide originally described as a dual-specificity protein phosphatase (2). The main substrates of PTEN are inositol phospholipids generated by the activation of the phosphoinositide 3-kinase (PI3K) (3). PTEN is a major negative regulator of the PI3K/Akt signaling pathway (1,4,5). PTEN possesses a carboxy-terminal, noncatalytic regulatory domain with three phosphorylation sites (Ser380, Thr382, and Thr383) that regulate PTEN stability and may affect its biological activity (6,7). PTEN regulates p53 protein levels and activity (8) and is involved in G protein-coupled signaling during chemotaxis (9,10). |
| 存放说明 | 4C |
| 参考文献 | Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5. Myers, M.P. et al. (1997) Proc Natl Acad Sci USA 94, 9052-7. Myers, M.P. et al. (1998) Proc Natl Acad Sci USA 95, 13513-8. Wan, X. and Helman, L.J. (2003) Oncogene 22, 8205-11. Wu, X. et al. (1998) Proc Natl Acad Sci USA 95, 15587-91. Vazquez, F. et al. (2000) Mol Cell Biol 20, 5010-8. Torres, J. and Pulido, R. (2001) J Biol Chem 276, 993-8. Freeman, D.J. et al. (2003) Cancer Cell 3, 117-30. Funamoto, S. et al. (2002) Cell 109, 611-23. Iijima, M. and Devreotes, P. (2002) Cell 109, 599-610. |
Figure 1. Phosphorylation of PTEN at Ser380 detected by PathScan® Phospho-PTEN (Ser380) Sandwich ELISA Kit #7285. PTEN in HeLa cells is constitutively phosphorylated at Ser380. Treatment of HeLa cell lysates with λ phosphatase (4000 U/mL for 60 minutes at 37ºC) abolishes the phosphorylation of PTEN at Ser380 as shown using PathScan® Phospho-PTEN (Ser380) Sandwich ELISA Kit and by Western analysis. The level of total PTEN detected by Western analysis remains unchanged. Absorbance450 nm is shown in the top figure while the corresponding Western blots using Phospho-PTEN (Ser380) Antibody #9551 (right panel) or PTEN Antibody #9552 (left panel) are shown in the bottom figure. Figure 1， 采用PathScan ® Phospho-PTEN (Ser380) Sandwich ELISA Kit #7285检测PTEN在Ser380位点的磷酸化 。PTEN在Hela细胞中Ser380位点的呈持续性磷酸化。 在λ 磷酸酶(4000 U/mL ，60 分钟 ，37°C)处理后，PathScan ® Phospho-PTEN (Ser380) Sandwich ELISA Kit 检测到PTEN在Ser380位点的磷酸化消失。Western 检测总PTEN蛋白未发生变化。在450nm处读取吸光度值（上图），相对应western blot采用抗体为Phospho-PTEN (Ser380) Antibody #9551 (右泳道) 或PTEN Antibody #9552（左泳道） ，见最下方图。 | |
