| 货号 | 7888C |
| 描述 | The PathScan® Total HER3/ErbB3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of HER3/ErbB3 protein. A HER3/ErbB3 mouse antibody has been coated on the microwells. After incubation with cell lysates, HER3/ErbB3 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a HER3/ErbB3 rabbit antibody is added to detect captured HER3/ErbB3 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 HER3/ErbB3 protein. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Total HER3/ErbB3 Sandwich ELISA Kit #7888 detects endogenous levels of total HER3/ErbB3 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 | HER3/ErbB3 is a member of the ErbB receptor protein tyrosine kinase family, but lacks tyrosine kinase activity. Tyrosine phosphorylation of ErbB3 depends on its association with other ErbB tyrosine kinases. Upon ligand binding, heterodimers form between ErbB3 and other ErbB proteins and ErbB3 is phosphorylated on tyrosine residues by the activated ErbB kinase (1,2). There are at least 9 potential tyrosine phosphorylation sites in the carboxy-terminal tail of ErbB3. These sites serve as consensus binding sites for signal transducing proteins, including Src family members, Grb2, and the p85 subunit of PI3 kinase, which mediate ErbB downstream signaling (3). Both Tyr1222 and Tyr1289 of ErbB3 reside within a YXXM motif and participate in signaling to PI3 kinase (4). ErbB3 is highly expressed in many cancer cells (5) and activation of the ErbB3/PI3K pathway is correlated with malignant phenotypes of adenocarcinomas (6). In tumor development, ErbB3 may function as an oncogenic unit together with other ErbB members (e.g. ErbB2 requires ErbB3 to drive breast tumor cell proliferation) (7). Thus, inhibiting interaction between ErbB3 and ErbB tyrosine kinases has become a novel strategy for anti-tumor therapy. |
| 存放说明 | 4C |
| 参考文献 | Yarden, Y. and Sliwkowski, M.X. (2001) Nat Rev Mol Cell Biol 2, 127-37. Guy, P.M. et al. (1994) Proc Natl Acad Sci U S A 91, 8132-6. Songyang, Z. et al. (1993) Cell 72, 767-78. Kim, H.H. et al. (1994) J Biol Chem 269, 24747-55. Sithanandam, G. et al. (2003) Carcinogenesis 24, 1581-92. Kobayashi, M. et al. (2003) Oncogene 22, 1294-301. Holbro, T. et al. (2003) Proc Natl Acad Sci U S A 100, 8933-8. |
Figure 1: Constitutive phosphorylation of HER3/ErbB3 in Calu-3 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit #7890 (top, right). In contrast, a low level of phospho-HER3/ErbB3 protein is detected in Calu-3 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total HER3/ErbB3 protein from both nonphospho or phospho lysates are detected by PathScan® Total HER3/ErbB3 Sandwich ELISA Kit #7888 (top, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-HER3/ErbB3 (Tyr1289) Rabbit mAb #4791 (right) or a total HER3/ErbB3 Antibody #4754 (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-HER3/ErbB3 (panTyr) Sandwich ELISA Kit #7890能够检测到 Calu-3细胞中HER3/ErbB3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-HER3/ErbB3的表达。用PathScan® Total HER3/ErbB3 Sandwich ELISA Kit #7888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HER3/ErbB3的表达（上，左）。上图显示450nm处的吸光值，下图显示用Phospho-HER3/ErbB3 (Tyr1289) Rabbit mAb #4791（右）或total HER3/ErbB3 Antibody #4754（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。 | |
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 the 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）。 |
