| 货号 | 7087C |
| 描述 | The PathScan® Phospho-TrkB (panTyr) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated TrkB protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A TrkB Mouse mAb 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 Biotinylated Phospho-Tyrosine Detection Antibody is added to detect captured tyrosine-phosphorylated TrkB protein. HRP-linked Streptavidin is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of tyrosine-phosphorylated TrkB 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-TrkB (panTyr) Chemiluminescent Sandwich ELISA Kit detects endogenous levels of TrkB protein when phosphorylated at Tyr residues in human cells. 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 (1). Neurotrophin signaling through these receptors regulates a number of physiological processes, such as cell survival, proliferation, neural development, and axon and dendrite growth and patterning (1). In the adult nervous system, the Trk receptors regulate synaptic strength and plasticity. TrkA regulates proliferation and is important for development and maturation of the nervous system (2). Phosphorylation at Tyr490 is required for Shc association and activation of the Ras-MAP kinase cascade (3,4). Residues Tyr674/675 lie within the catalytic domain, and phosphorylation at these sites reflects TrkA kinase activity (3-6). Point mutations, deletions, and chromosomal rearrangements (chimeras) cause ligand-independent receptor dimerization and activation of TrkA (7-10). TrkA is activated in many malignancies including breast, ovarian, prostate, and thyroid carcinomas (8-13). Research studies suggest that expression of TrkA in neuroblastomas may be a good prognostic marker as TrkA signals growth arrest and differentiation of cells originating from the neural crest (10).Many tyrosine phosphorylation sites are conserved between TrkA and TrkB: Tyr490 of TrkA corresponds to Tyr512 in TrkB, and Tyr674/675 of TrkA to Tyr706/707 in TrkB of the human sequence (14). TrkB is overexpressed in tumors such as neuroblastoma, prostate adenocarcinoma, and pancreatic ductal adenocarcinoma (15). Research studies have shown that in neuroblastomas, overexpression of TrkB correlates with an unfavorable disease outcome when autocrine loops signaling tumor survival are potentiated by additional overexpression of brain-derived neurotrophic factor (BDNF) (16-18). An alternatively spliced truncated TrkB isoform lacking the kinase domain is overexpressed in Wilms’ tumors and this isoform may act as a dominant-negative regulator of TrkB signaling (17). |
| 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. Relationship between protein concentration of lysates from untreated and BDNF-treated 3T3/TrkB cells and immediate light generation with chemiluminescent substrate is shown. After starvation, 3T3/TrkB cells (85% confluent) were treated with Human Brain-Derived Neurotrophic Factor (BDNF) #3897 (100 ng/ml, 2 min) at 37°C, and lysed with 1X Cell Lysis Buffer (10X) #9803. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range. |
