| 货号 | 7111C |
| 描述 | The PathScan® Phospho-TrkB (Tyr516) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected levels of TrkB when phosphorylated at Tyr516. A TrkB mouse antibody has been coated onto the microwells. After incubation with cell lysates, TrkB (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-TrkB (Tyr516) rabbit detection antibody is added to detect tyrosine phosphorylation of the captured TrkB 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 TrkB phosphorylated on Tyr516. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-TrkB (Tyr516) Sandwich ELISA Kit #7111 detects transfected levels of TrkB protein when Tyr516 is phosphorylated (see 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 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. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42. Geiger, T.R. and Peeper, D.S. (2005) Cancer Res 65, 7033-6. Han, L. et al. (2007) Med Hypotheses 68, 407-9. Aoyama, M. et al. (2001) Cancer Lett 164, 51-60. Desmet, C.J. and Peeper, D.S. (2006) Cell Mol Life Sci 63, 755-9. |
Figure 1: Treatment of 3T3/TrkB cells with BDNF stimulates tyrosine-phosphorylation of TrkB, detected by PathScan® Phospho-TrkB (Tyr516) Sandwich ELISA Kit #7111, but does not affect the levels of total TrkB detected by PathScan® Total TrkB Sandwich ELISA Kit #7106. Absorbance at 450 nm is shown in the top figure, while the corresponding Western blots using Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9) Rabbit mAb #4619 (right panel) or TrkB (80E3) Rabbit mAb #4603 (left panel), are shown in the bottom figure. Human TrkB is transfected and expressed in NIH/3T3 cells. 图1：NGF处理后的3T3/TrkB细胞产生了TrkB的酪氨酸磷酸化，为PathScan Phospho-TrkB (Tyr516) Sandwich ELISA kit #7111所检测到，但是却并不产生PathScan Total TrkB Sandwich ELISA kit #7106可检测到的总TrkB水平的变化。450nm吸光读数在上图显示，对应的使用Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9)Rabbit mAb 兔单抗#4619（右）或TrkB (80E3)Rabbit mAb 兔单抗#4603（左）的结果在下图显示。人TrkB转染至NIH/3T3细胞中表达。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and BDNF-treated 3T3/TrkB cells and the absorbance at 450 nm is shown. 3T3/TrkB cells (85% confluence) were starved and treated with BDNF (100 ng/ml) for 2 min at 37ºC, and then lysed. 图2：显示未处理和经BDNF处理的3T3/TrkB细胞提取物中的蛋白浓度与试剂盒光学浓度读数的关系。3T3/TrkB细胞(85% confluence)用100ng/ml的BDNF在37度处理两分钟，然后裂解。 |
