| 货号 | 7310C |
| 描述 | The PathScan® Total HER2/ErbB2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of 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 HER2/ErbB2 rabbit antibody is added to detect captured 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. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Total HER2/ErbB2 Sandwich ELISA Kit #7310 detects endogenous levels of total HER2/ErbB2 protein in human cells, as shown in 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. |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | 4C |
| 参考文献 | Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9. |
Figure 2: The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. Unstarved Calu-3 cells were cultured (85% confluence) and lysed with or without addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate).图2：磷酸化或非磷酸化的裂解中蛋白含量和450nm吸光值之间的关系。未饥饿的Calu-3细胞在85%满时收样，并用加入或未加磷酸酶抑制剂的裂解液进行处理（磷酸化或非磷酸化）。 | |
Figure 1: Constitutive phosphorylation of HER2/ErbB2 in Calu-3 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-HER2/ErbB2 (panTyr) Sandwich ELISA Kit #7968 (top, right). In contrast, a low level of phospho-HER2/ErbB2 protein is detected in Calu-3 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total HER2/ErbB2 protein from both nonphospho or phospho lysates are detected by PathScan® Total HER2/ErbB2 Sandwich ELISA Kit #7310 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-HER2/ErbB2 (Tyr1221/1222) Rabbit mAb #2243 (right) or a total HER2/ErbB2 Antibody #2242 (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4. 图1：在磷酸酶抑制剂*存在时（磷酸化的裂解液），用PathScan® Phospho-HER2/ErbB2 (panTyr) Sandwich ELISA Kit #7968能够检测到Calu-3细胞中phospho-HER2/ErbB2的组成型表达（上，右）。相比之下，没有磷酸酶抑制剂*存在时（非磷酸化的裂解液），仅能检测到低水平的phospho-HER2/ErbB2的表达。用Scan® Total HER2/ErbB2 Sandwich ELISA Kit #7310则能在磷酸化和非磷酸化的裂解液中检测到近似水平的总HER2/ErbB2的表达（上，左）。上图显示450nm处的吸光值，下图显示用Phospho-HER2/ErbB2 (Tyr1221/1222) Rabbit mAb #2243（右）或total HER2/ErbB2 Antibody #2242（左）进行免疫印迹检测的结果。*磷酸酶抑制剂中包含sodium pyrophosphate、β-glycerophosphate和 Na3VO4。 |
