| 货号 | 7034C |
| 描述 | The PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Ret protein. A Ret rabbit mAb has been coated on the microwells. After incubation with cell lysates, Ret protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse mAb is added to detect captured tyrosine-phosphorylated Ret 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 Ret protein phosphorylated on tyrosine residues. Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit #7034 detects endogenous levels of tyrosine-phosphorylated Ret protein 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 Ret proto-oncogene (c-Ret) is a receptor tyrosine kinase that functions as a multicompetent receptor complex in conjunction with other membrane-bound ligand-binding GDNF family receptors (1). Ligands that bind the Ret receptor include the glial cell line-derived neurotropic factor (GDNF) and its congeners neurturin, persephin and artemin (2-4). Alterations in the corresponding Ret gene are associated with diseases including papillary thyroid carcinoma, multiple endocrine neoplasia (type 2A and 2B), familial medullary thyroid carcinoma and a congenital developmental disorder known as Hirschsprung’s disease (1,3). The Tyr905 residue located in the Ret kinase domain plays a crucial role in Ret catalytic and biological activity. Substitution of Phe for Tyr905 dramatically inhibits Ret autophosphorylation activity (5). |
| 存放说明 | 4C |
| 参考文献 | Airaksinen, M.S. et al. (1999) Mol Cell Neurosci 13, 313-25. Takahashi, M. et al. (1989) Oncogene 4, 805-6. Manié, S. et al. (2001) Trends Genet 17, 580-9. Tallini, G. and Asa, S.L. (2001) Adv Anat Pathol 8, 345-54. Iwashita, T. et al. (1999) Oncogene 18, 3919-22. |
Figure 1: Constitutive phosphorylation of Ret in TT cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit #7034 (upper, right). In contrast, a low level of phospho-Ret protein is detected in TT cells lysed in the absence of phosphatase inhibitors* (nonphospho lysate). Similar levels of total Ret protein from both nonphospho or phospho lysates are detected by PathScan® Total Ret Sandwich ELISA Kit #7032 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using a Phospho-Ret (Tyr905) Rabbit Antibody #3221 (right) or a total Ret (C31B4) Rabbit mAb #3223 (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-Ret (panTyr) Sandwich ELISA Kit #7034能够检测到NTT细胞中磷酸化Ret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磷酸化Ret的表达。用PathScan® Total Ret Sandwich ELISA Kit #7032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Ret的表达（上，左）。上图显示450nm处的吸光值，下图显示用Phospho-Ret (Tyr905) Rabbit Antibody #3221（右）或total Ret (C31B4) Rabbit mAb #3223（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. TT 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。TT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）。 |
