PathScan? Phospho-TrkB (Tyr516) Sandwich ELISA Kit

• 产品编号：CST-7111C      品牌：CST       原厂货号：7111C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

PathScan®Phospho-TrkB (Tyr516) Sandwich ELISA Kit #7111可识别内源性的Tyr516磷酸化的TrkB，如图1所示。试剂盒敏感性如图2所示。本试剂盒可检测经内部测试确定的指定物种的蛋白质，同时也可以检测其他物种的同源蛋白。PathScan®Phospho-TrkB (Tyr516) Sandwich ELISA Kit是固相夹心酶联免疫吸附试验（ELISA）试剂盒，识别转染水平的Tyr516磷酸化的TrkB。TrkB鼠抗体已经被包被在微孔上。在和细胞裂解液孵育时，TrkB蛋白（磷酸化和非磷酸化）将被包被抗体捕获。在接下来的洗涤后，Tyr516磷酸化的TrkB鼠检测抗体将加入，以便识别捕获的酪氨酸磷酸化的TrkB蛋白。抗兔IgG，HRP-偶联的IgG抗体，将用来识别连接的检测抗体。HRP底物TMB将用来显色。显色后的吸光程度正比于Tyr516磷酸化的TrkB的量。Trk受体酪氨酸激酶家族包括TrkA，TrkB和TrkC。这些家庭成员的序列是高度保守的，它们是由不同的神经营养因子激活：NGF激活TrkA，BDNF或NT4激活TrkB，NT3激活TrkC。TrkA调节细胞增殖，神经系统的发育和成熟很重要（1）。在Tyr490磷酸化，需要与Shc结合及Ras-MAP激酶级联的激活。Tyr674/675残基在催化域内，此位点的磷酸化能影响TrkA的激酶活性（2-6）。点突变，缺失和染色体重排（嵌合体）会引起配体依赖受体的二聚化，激活TrkA。许多恶性肿瘤，包括乳腺癌、结肠癌、前列腺癌、甲状腺癌和急性髓细胞白血病有激活的TrkA。神经母细胞瘤的TrkA表达是一个预后良好的指标，因为它标志着肿瘤生长停滞和从神经嵴分化而来（1）。

1. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
2. Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89.
3. Stephens, R.M. et al. (1994) Neuron 12, 691-705.
4. Marsh, H.N. et al. (2003) J Cell Biol 163, 999-1010.
5. Obermeier, A. et al. (1993) EMBO J 12, 933-41.
6. Obermeier, A. et al. (1994) EMBO J 13, 1585-90.
7. Arevalo, J.C. et al. (2001) Oncogene 20, 1229-34.
8. Reuther, G.W. et al. (2000) Mol Cell Biol 20, 8655-66.
9. Greco, A. et al. (1997) Genes Chromosomes Cancer 19, 112-23.
10. Pierotti, M.A. and Greco, A. (2006) Cancer Lett 232, 90-8.
11. Lagadec, C. et al. (2009) Oncogene 28, 1960-70.
12. Greco, A. et al. (2010) Mol Cell Endocrinol 321, 44-9.
13. Ødegaard, E. et al. (2007) Hum Pathol 38, 140-6.
14. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
15. Geiger, T.R. and Peeper, D.S. (2005) Cancer Res 65, 7033-6.
16. Han, L. et al. (2007) Med Hypotheses 68, 407-9.
17. Aoyama, M. et al. (2001) Cancer Lett 164, 51-60.
18. Desmet, C.J. and Peeper, D.S. (2006) Cell Mol Life Sci 63, 755-9.

原厂资料：

Specificity / Sensitivity

CST's 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.

Description

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.

Background

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).

1. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
2. Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89.
3. Stephens, R.M. et al. (1994) Neuron 12, 691-705.
4. Marsh, H.N. et al. (2003) J Cell Biol 163, 999-1010.
5. Obermeier, A. et al. (1993) EMBO J 12, 933-41.
6. Obermeier, A. et al. (1994) EMBO J 13, 1585-90.
7. Arevalo, J.C. et al. (2001) Oncogene 20, 1229-34.
8. Reuther, G.W. et al. (2000) Mol Cell Biol 20, 8655-66.
9. Greco, A. et al. (1997) Genes Chromosomes Cancer 19, 112-23.
10. Pierotti, M.A. and Greco, A. (2006) Cancer Lett 232, 90-8.
11. Lagadec, C. et al. (2009) Oncogene 28, 1960-70.
12. Greco, A. et al. (2010) Mol Cell Endocrinol 321, 44-9.
13. Ødegaard, E. et al. (2007) Hum Pathol 38, 140-6.
14. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
15. Geiger, T.R. and Peeper, D.S. (2005) Cancer Res 65, 7033-6.
16. Han, L. et al. (2007) Med Hypotheses 68, 407-9.
17. Aoyama, M. et al. (2001) Cancer Lett 164, 51-60.
18. Desmet, C.J. and Peeper, D.S. (2006) Cell Mol Life Sci 63, 755-9.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).