PathScan? Phospho-HER2/ErbB2 (panTyr) Sandwich ELISA Kit

• 产品编号：CST-7968C      品牌：CST       原厂货号：7968C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

CST公司的PathScan® Phospho-HER2/ErbB2 (panTyr) Sandwich ELISA Kit #7968用于检测内源酪氨酸位点磷酸化的HER2/ErbB2蛋白的表达（如图１）。图2显示试剂盒的灵敏度。本试剂盒不与其它ErbB家族的蛋白发生交叉反应（数据未显示）。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-HER2/ErbB2 (panTyr) Sandwich ELISA Kit利用固相双抗夹心酶联免疫检测法对内源性酪氨酸磷酸化HER2/ErbB2的表达进行检测。微孔中已经包被了抗体HER2/ErbB2鼠抗体，与细胞裂解液共孵育后，磷酸化的和未磷酸化的HER2/Erb2受体蛋白均能被包被抗体捕获。多次洗板之后，加入磷酸化酪氨酸兔检测抗体对被捕获的酪氨酸磷酸化HER2/ErbB2蛋白进行检测，然后加入抗兔IgG, HRP偶联抗体对检测抗体进行识别，最后加入HRP的底物TMB进行显色反应。按照相对光单位，发射光强度与酪氨酸位点磷酸化的HER2/ErbB2蛋白的量成正比。原癌基因ErbB2 (HER2)编码185kDa的受体样跨膜糖蛋白，该蛋白具有固有的酪氨酸激酶的活性 (1)。ErbB2的配体目前尚不清楚，但在高表达的ErbB2与其它ErbB家族成员形成异源二聚体时，即使缺少配体，其酶活性仍能被激活(2)。将近40%的人类乳腺癌中能够检测到ErbB2基因的扩增及其产物的过量表达。泛素连接酶c-Cbl结合在ErbB2的Tyr1112上能够导致ErbB2聚泛素化并促进其降解(4)。ErbB2是乳腺癌等多种癌肿的关键治疗靶点，通过c-Cbl介导的蛋白水解途径靶向调控ErbB2的降解是一种潜在的治疗手段。磷酸化ErbB2的激酶位点Tyr877（同源于pp60c-Src 的Tyr416）能调控ErbB2的生物活性。ErbB2的自磷酸化位点是Tyr1248和 Tyr1221/1222，这些位点的磷酸化将ErbB2与Ras-Raf-MAP激酶的信号转导途径联系起来(1,5)。

1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-HER2/ErbB2 (panTyr) Sandwich ELISA Kit #7968 detects endogenous levels of HER2/ErbB2 protein only when phosphorylated at Tyr residues (see Figure 1). The kit sensitivity is shown in Figure 2. This kit does not cross-react with other proteins of ErbB family (data not shown). 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-HER2/ErbB2 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated HER2/ErbB2 protein. A HER2/ErbB2 mouse antibody has been coated on the microwells. After incubation with cell lysates, HER2/ErbB2 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine rabbit detection antibody is added to detect captured tyrosine-phosphorylated HER2/ErbB2 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 HER2/ErbB2 protein phosphorylated on tyrosine.

Background

The ErbB2 (HER2) proto-oncogene encodes a 185 kDa transmembrane, receptor-like glycoprotein with intrinsic tyrosine kinase activity (1). While ErbB2 lacks an identified ligand, ErbB2 kinase activity can be activated in the absence of a ligand when overexpressed and through heteromeric associations with other ErbB family members (2). Amplification of the ErbB2 gene and overexpression of its product are detected in almost 40% of human breast cancers (3). Binding of the c-Cbl ubiquitin ligase to ErbB2 at Tyr1112 leads to ErbB2 poly-ubiquitination and enhances degradation of this kinase (4). ErbB2 is a key therapeutic target in the treatment of breast cancer and other carcinomas and targeting the regulation of ErbB2 degradation by the c-Cbl-regulated proteolytic pathway is one potential therapeutic strategy. Phosphorylation of the kinase domain residue Tyr877 of ErbB2 (homologous to Tyr416 of pp60c-Src) may be involved in regulating ErbB2 biological activity. The major autophosphorylation sites in ErbB2 are Tyr1248 and Tyr1221/1222; phosphorylation of these sites couples ErbB2 to the Ras-Raf-MAP kinase signal transduction pathway (1,5).

1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.

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).