| 货号 | 12737C |
| 描述 | PathScan® Phospho-Ron (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Ron proteins. A Ron Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Ron protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, Phospho-Tyrosine Mouse Detection Antibody is added to detect captured tyrosine-phosphorylated Ron 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 absorbance for the developed color is proportional to the quantity of tyrosine-phosphorylated Ron protein. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-Ron (panTyr) Sandwich ELISA Kit detects endogenous levels of Ron protein when 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 | Ron is a member of the Met protooncogene family of receptor tyrosine kinases, which also includes Stk, c-Met, and c-Sea. The functional Ron is a heterodimer composed of a 40 kDa α chain and a 150 kDa β chain. Ron is initially synthesized in the cells as a single-chain, pro-Ron precursor that is cleaved into the two active chains. The α chain is completely extracellular, whereas the β chain traverses the cell membrane and contains the intracellular tyrosine kinase and regulatory elements (1,2). Ron mediates multiple signaling cascades that involve cell motility, adhesion, proliferation, and apoptosis. The signaling pathways activated downstream of Ron include the ras/mitogen-activated protein kinase (MAPK), phosphatidyl inositol-3 kinase (PI3K)/Akt, and focal adhesion kinase (FAK) pathways. Ron activation can also significantly increase c-Src activity, a signaling intermediate involved in cell cycle progression, motility, angiogenesis and survival (3,4). The function of Ron has been shown to be important for embryological development as well as implicated in the progression and metastasis of tumors (5). |
| 存放说明 | 4C |
| 参考文献 | Ronsin, C. et al. (1993) Oncogene 8, 1195-202. |
Figure 1. Constitutive phosphorylation of Ron in HCC827 cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-Ron (panTyr) Sandwich ELISA Kit. In contrast, a low level of phospho-Ron protein is detected in HCC827 cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Ron (81H9) Rabbit mAb #2654 (left) and a Phospho-Ron (Tyr1238/1239) antibody (right) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and sodium orthovanadate.图1.在存在磷酸酶抑制剂*（磷酸化裂解物）的情况下，HCC827细胞裂解物中Ron的组成型磷酸化用PathScan® Phospho-Ron (panTyr) Sandwich ELISA Kit进行检测。相反，在缺少磷酸酶抑制剂*（非磷酸化裂解物）的情况下，检测HCC827细胞裂解物中低水平的磷酸化Ron蛋白。上图显示450nm处的吸光度值，而对应的western blot实验见下图，western blot实验使用的抗体为Ron (81H9) Rabbit mAb #2654 (左图)和 Phospho-Ron (Tyr1238/1239) antibody (右图)。*磷酸酶抑制剂包括焦磷酸钠，β-甘油磷酸和原钒酸钠。 | |
Figure 2. The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. Unstarved HCC827 cells were cultured (85% confluence) and lysed with or without addition of phosphatase inhibitors to the lysis buffer (phospho or nonphospho lysate, respectively).图2.如图所示，裂解物中磷酸化和非磷酸化蛋白浓度与450nm处吸光度值的关系。未饥饿处理HCC827细胞长至85%汇合，用或不用添加磷酸酶抑制剂的裂解物缓冲液（分别为磷酸化和非磷酸化裂解物）进行裂解。 |
