PathScan? Phospho-Met (panTyr) Sandwich ELISA Antibody Pair

• 产品编号：CST-7334S      品牌：Cell Signaling Technology       原厂货号：7334S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测抗体对
• 应用分类：

其它组分：

描述：

抗体对的特异性和敏感性请参考相应的PathScan® Sandwich ELISA Kit。注意：内部检测显示，本试剂盒能检测列举出的物种蛋白，但不排除用于其它物种同源蛋白检测的可能。CST公司的PathScan® Phospho-Met (panTyr) Sandwich ELISA Antibody Pair是PathScan® Phospho-Met (panTyr) Sandwich ELISA Kit #7333的另一种经济型选择，主要提供捕获和检测抗体（100 X贮存液）以及HRP偶联二抗 (1000X 贮存液），可供4 x 96孔ELISA检测使用。用PBS稀释后的Met兔捕获抗体包被96孔板，封闭后加入细胞裂解液孵育，然后加入酪氨酸磷酸化鼠检测抗体和HRP偶联二抗。最后加入HRP的底物TMB进行显色反应。按照相对光单位，发射光强度与磷酸化Met (panTyr)蛋白的量成正比。*试剂盒中的抗体为试剂盒特异定制配方。Met作为肝细胞生长因子 (HGF, 亦作扩散因子) 的高亲和性酪氨酸激酶受体是一个二硫键连接的异二聚体，含有一个45 kDa的 α-亚基和一个145 kDa的 β-亚基 (1,2)。α-亚基和β-亚基的氨基端形成胞外区，β-亚基的其余部分跨细胞膜并且含有一个具有激酶活性的胞内区。Met和HGF相互作用会导致多个酪氨酸位点的自磷酸化，继而招募下游信号分子Gab1, c-Cbl和 PI3 K激酶（3）。这一系列基础性事件对于Met激酶相关的所有生物学功能十分重要。胞内Tyr1003位点的磷酸化对于Met的泛素化和降解是必须的（4）。激酶结构域Tyr1234/1235的磷酸化对于Met的激酶活性十分关键。胞内 Tyr1349的磷酸化为Met提供了Gab1 的直接结合位点(5)。在肾、结肠和乳腺癌中发现有Met水平和/或激酶活性的改变，因此Met是肿瘤治疗和诊断的一个靶点（6,7）。

1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

原厂资料：

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Phospho-Met (panTyr) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-Met (panTyr) Sandwich ELISA Kit #7333. Capture and Detection Antibodies (100X stocks) and HRP-Conjugated Secondary Antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Met Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by Phospho-Tyrosine Mouse Detection Antibody and HRP-conjugated Secondary Antibody. HRP substrate (TMB) is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Met (panTyr) protein. 
 *Antibodies in this kit are custom formulations specific to the kit.

Background

Met, a high affinity tyrosine kinase receptor for hepatocyte growth factor (HGF, also known as scatter factor) is a disulfide-linked heterodimer made of 45 kDa α- and 145 kDa β-subunits (1,2). The α-subunit and the amino-terminal region of the β-subunit form the extracellular domain. The remainder of the β-chain spans the plasma membrane and contains a cytoplasmic region with tyrosine kinase activity. Interaction of Met with HGF results in autophosphorylation at multiple tyrosines, which recruit several downstream signaling components, including Gab1, c-Cbl, and PI3 kinase (3). These fundamental events are important for all of the biological functions involving Met kinase activity. The addition of a phosphate at cytoplasmic Tyr1003 is essential for Met protein ubiquitination and degradation (4). Phosphorylation at Tyr1234/1235 in the Met kinase domain is critical for kinase activation. Phosphorylation at Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon, and breast. Thus, Met is an attractive cancer therapeutic and diagnostic target (6,7).

1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

注意事项：

Storage: Capture and Detection Antibodies are stored at
4ºC. HRP-conjugated Secondary Antibody is stored at -20ºC.