| 货号 | 7093C |
| 描述 | CSTs PathScan® Phospho-Ros (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Ros protein. A Ros rabbit antibody has been coated on the microwells. After incubation with cell lysates, Ros protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse detection antibody is added to detect captured tyrosine-phosphorylated Ros protein. 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 Ros protein phosphorylated on tyrosine. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-Ros (panTyr) Sandwich ELISA Kit #7093 detects endogenous levels of Ros protein phosphorylated at Tyr residues in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. 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 c-ros proto-oncogene encodes a receptor tyrosine kinase similar to insulin receptor proteins and expressed in specific epithelial cells (1). During mouse development the Ros receptor plays an important role in the differentiation of epididymal epithelium (2). Overexpression or mutation of Ros may lead to oncogenic development. Furthermore, stable transfection with a chimeric receptor containing the Ros transmembrane and cytoplasmic domains leads to transformation of NIH/3T3 fibroblasts (3). A genetic rearrangement that produces a fusion protein containing Ros and FIG (fused in glioblastoma) results in glioblastoma formation (4). The FIG-Ros fusion abnormally activates the SHP-2 phosphatase and PI3K/Akt/mTOR pathways in tumor cells (5). Additional Ros mutations are associated with NSCLC cancer cells indicating that Ros may be a candidate for tumor targeted therapy (6). |
| 存放说明 | 4C |
| 参考文献 | Matsushime, H. et al. (1986) Mol Cell Biol 6, 3000-4. Yeung, C.H. et al. (1999) Biol Reprod 61, 1062-9. Charest, A. et al. (2003) Genes Chromosomes Cancer 37, 58-71. Gu, T.L. et al. (2011) PLoS One 6, e15640. Birch, A.H. et al. (2011) PLoS One 6, e28250. Rimkunas, V.M. et al. (2012) Clin Cancer Res 18, 4449-57. Rikova, K. et al. (2007) Cell 131, 1190-203. Stumpfova, M. and Jänne, P.A. (2012) Clin Cancer Res 18, 4222-4. Jun, H.J. et al. (2012) Cancer Res 72, 3764-74. |
Figure 1: Constitutive phosphorylation of Ros in HCC78 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-Ros (panTyr) Sandwich ELISA Kit #7093 (upper, right). In contrast, a low level of phospho-Ros protein is detected in HCC78 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total Ros protein from both nonphospho and phospho lysates are detected by PathScan® Total Ros Sandwich ELISA Kit #7091 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a Phospho-Ros (Tyr2274) Antibody #3078 (right) or a total Ros Mouse mAb #3266 (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-Ros (panTyr) Sandwich ELISA Kit #7093能够检测到HCC78细胞中磷酸化Ros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HCC78细胞中低水平的磷酸化Ros的表达。用PathScan® Total Ros Sandwich ELISA Kit #7091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Ros的表达（上，左）。上图显示450nm处的吸光值，下图显示用Phospho-Ros (Tyr2274) Antibody #3078（右）或total Ros (69D6) Mouse mAb #3266（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. HCC78 cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate, respectively).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。HCC78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）。 |
