PathScan? Total PTEN Sandwich ELISA Kit

• 产品编号：CST-7882C      品牌：CST       原厂货号：7882C
• 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 PTEN Sandwich ELISA Kit #7882可以检测内源性的PTEN蛋白，如图1所示， 采用Total PTEN Sandwich ELISA Kit #7882可以在PTEN阳性的细胞系中检测到高水平的PTEN，而在PTEN阴性细胞PC-3中则无法检测到。本试剂盒经过内部测试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 PTEN 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PTEN蛋白。兔源PTEN抗体已经包被在孔中，与细胞的裂解液共孵育后， PTEN蛋白被已包被的抗体捕获。经过彻底的洗涤，加入鼠源PTEN检测抗体去检测被捕获PTEN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后产生颜色。颜色的吸光度值与PTEN蛋白的含量呈正相关。PTEN（phosphatase and tensin homologue deleted on chromosome ten），也称为MMAC（ (mutated in multiple advanced cancers)磷酸酶，是一种存在于多种人类癌症中的肿瘤抑制因子（1）。PTEN编码一个403个氨基酸组成的多肽，最初被认为是一个双特异性蛋白磷酸酶（2）。PTEN的主要底物是3磷酸肌醇激酶（PI3K）激活转化生成的肌醇磷脂（3）。PTEN是PI3K/Akt信号通路的一个主要的负调因子（1,4,5）。PTEN有一个羧基端的非催化调控的结构域，其中含有3个可以调节PTEN的稳定性，也可能影响其生物活性的磷酸化位点（Ser380，Thr382，Thr383）（6,7）。PTEN调节p53蛋白水平和活性（8），并在趋化过程中参与G蛋白偶联信号转导（9,10）。

1. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
2. Myers, M.P. et al. (1997) Proc Natl Acad Sci USA 94, 9052-7.
3. Myers, M.P. et al. (1998) Proc Natl Acad Sci USA 95, 13513-8.
4. Wan, X. and Helman, L.J. (2003) Oncogene 22, 8205-11.
5. Wu, X. et al. (1998) Proc Natl Acad Sci USA 95, 15587-91.
6. Vazquez, F. et al. (2000) Mol Cell Biol 20, 5010-8.
7. Torres, J. and Pulido, R. (2001) J Biol Chem 276, 993-8.
8. Freeman, D.J. et al. (2003) Cancer Cell 3, 117-30.
9. Funamoto, S. et al. (2002) Cell 109, 611-23.
10. Iijima, M. and Devreotes, P. (2002) Cell 109, 599-610.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Total PTEN Sandwich ELISA Kit #7882 detects endogenous levels of PTEN. As shown in Figure 1, using the Total PTEN Sandwich ELISA Kit #7882, a high level of PTEN is detected in a variety of PTEN positive cell lines but not in the PTEN negative cell line PC-3. 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 PTEN Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenously expressed PTEN. A PTEN rabbit antibody has been coated onto the microwells. After incubation with cell lysates, PTEN is captured by the coated antibody. Following extensive washing, a PTEN mouse detection antibody is added to detect the captured PTEN. 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 PTEN.

Background

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).

1. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
2. Myers, M.P. et al. (1997) Proc Natl Acad Sci USA 94, 9052-7.
3. Myers, M.P. et al. (1998) Proc Natl Acad Sci USA 95, 13513-8.
4. Wan, X. and Helman, L.J. (2003) Oncogene 22, 8205-11.
5. Wu, X. et al. (1998) Proc Natl Acad Sci USA 95, 15587-91.
6. Vazquez, F. et al. (2000) Mol Cell Biol 20, 5010-8.
7. Torres, J. and Pulido, R. (2001) J Biol Chem 276, 993-8.
8. Freeman, D.J. et al. (2003) Cancer Cell 3, 117-30.
9. Funamoto, S. et al. (2002) Cell 109, 611-23.
10. Iijima, M. and Devreotes, P. (2002) Cell 109, 599-610.

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).