| 货号 | 7210C |
| 描述 | PathScan® Phospho-TrkA (Tyr490) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects phospho-TrkA (Tyr490) protein. A TrkA Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-TrkA proteins are captured by the coated antibody. Following extensive washing, Phospho-TrkA (Tyr490) Antibody is added to detect phospho-TrkA 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 optical density for this developed color is proportional to the quantity of Phospho-TrkA (Tyr490) 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-TrkA (Tyr490) Sandwich ELISA Kit #7210 detects phospho-TrkA (Tyr490) Protein. As shown in Figure 1, using this ELISA Kit #7210, a significant induction of Phospho-TrkA (Tyr490) is detected in 3T3/TrkA cells treated with NGF. However, levels of total TrkA (phospho and nonphospho) detected by PathScan® Total TrkA Sandwich ELISA Kit #7208, remain unchanged (Figure 1). 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 family of Trk receptor tyrosine kinases consists of TrkA, TrkB and TrkC. While the sequence of these family members is highly conserved, they are activated by different neurotrophins: TrkA by NGF, TrkB by BDNF or NT4, and TrkC by NT3. TrkA regulates proliferation and is important for development and maturation of the nervous system (1). Phosphorylation at Tyr490 is required for Shc association and activation of the Ras-MAP kinase cascade. Residues Tyr674/675 lie within the catalytic domain, and phosphorylation at this site reflects TrkA kinase activity (2-6). Point mutations, deletions and chromosomal rearrangements (chimeras) cause ligand-independent receptor dimerization and activation of TrkA. Many malignancies including breast, colon, prostate and thyroid carcinomas and acute myeloid leukemia have activated TrkA. Expression of TrkA in neuroblastomas is a good prognostic marker because it signals growth arrest and differentiation of cells originating from the neural crest (1). |
| 存放说明 | 4C |
| 参考文献 | Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42. Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89. Stephens, R.M. et al. (1994) Neuron 12, 691-705. Marsh, H.N. et al. (2003) J Cell Biol 163, 999-1010. Obermeier, A. et al. (1993) EMBO J 12, 933-41. Obermeier, A. et al. (1994) EMBO J 13, 1585-90. Arevalo, J.C. et al. (2001) Oncogene 20, 1229-34. Reuther, G.W. et al. (2000) Mol Cell Biol 20, 8655-66. Greco, A. et al. (1997) Genes Chromosomes Cancer 19, 112-23. Pierotti, M.A. and Greco, A. (2006) Cancer Lett 232, 90-8. Lagadec, C. et al. (2009) Oncogene 28, 1960-70. Greco, A. et al. (2010) Mol Cell Endocrinol 321, 44-9. Ødegaard, E. et al. (2007) Hum Pathol 38, 140-6. |
Figure 1: Treatment of 3T3/TrkA cells with NGF stimulates phosphorylation of TrkA at Tyr490, detected by PathScan® Phospho-TrkA (Tyr490) Sandwich ELISA Kit #7210, but does not affect the level of total TrkA detected by PathScan® Total TrkA Sandwich ELISA Kit #7208. OD 450 readings are shown in the top figure, while the corresponding western blot using Phospho-TrkA (Tyr490) Antibody #9141 (right panel) or TrkA Rabbit mAb #2505 (left panel), is shown in the bottom figure. The human TrkA is expressed in 3T3/TrkA cells. 图1：NGF处理后的3T3/TrkA细胞产生了TrkA的Tyr490磷酸化，为PathScan Phospho-TrkA (Tyr490) Sandwich ELISA kit #7210所检测到，但是却并不产生PathScan Total TrkA Sandwich ELISA kit #7208可检测到的总TrkA水平的变化。OD 450 读数在上图显示，对应的使用Phospho-TrkA (Tyr490) Antibody 兔多抗 #9141（右）或TrkA Rabbit mAb 兔单抗 #2505（左）的结果在下图显示。人TrkA在3T3/TrkA细胞中表达。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and NGF-treated 3T3/TrkA cells and kit assay optical density readings is shown. After starvation, 3T3/TrkA cells (85% confluence) were treated with NGF (100 ng/ml) for 2 min at 37°C, and then lysed. 图2：显示未处理和经NGF处理的3T3/TrkA细胞提取物中的蛋白浓度与试剂盒光学浓度读数的关系。饥饿处理后，3T3/TrkA细胞(85% confluence)用100ng/ml的NGF在37度处理两分钟，然后裂解。 |
