PathScan? Phospho-TrkA (Tyr490) Chemiluminescent Sandwich ELISA Kit

• 产品编号：CST-7024C      品牌：CST       原厂货号：7024C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

PathScan® Phospho-TrkA (Tyr490) Chemiluminescent Sandwich ELISA Kit #7024 能够检测人细胞中内源性的磷酸化TrkA(Tyr490)。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-TrkA (Tyr490) Chemiluminescent Sandwich ELISA Kit采用了固相夹心酶联免疫吸附方法（ELISA），通过化学发光读数检测内源性的TrkA蛋白Tyr490位点磷酸化的情况。与传统的显色检测法相比，化学发光ELISA 试剂盒往往有着更宽泛的动态范围和更高的灵敏度。这种化学发光ELISA采用低体积的微孔板，在加强信号和灵敏度的同时需要的样本量更少。微孔中包被了TrkA Mouse mAb鼠单抗。细胞裂解液孵育后，磷酸化和非磷酸化TrkA蛋白被包被的抗体捕获。充分洗涤后，添加Phospho-TrkA (Tyr490) Rabbit Detection Antibody检测捕获的TrkA蛋白。再用Anti- rabbit IgG, HRP-linked Antibody识别结合的抗体。添加化学发光试剂增强信号。测得的相对光单位（RLU）与磷酸化TrkA（Tyr490）蛋白的量成正比。试剂盒中的抗体为该试剂盒所特制。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（1）。TrkA调节细胞增殖，神经系统的发育和成熟很重要（1）。神经营养因子信号通过这些受体调节了一些生理过程，如细胞的生存，增殖，神经系统的发育，轴突和枝晶的生长和构成（1）。在成年神经系统，Trk受体调节突触的强度和塑性。 TrkA调节细胞增殖，它对于神经系统的发育和成熟是非常重要的（2）。在Tyr490磷酸化，需要Shc的结合及Ras-MAP激酶级联的激活（3,4）。Tyr674/675残基在催化域内，这些位点的磷酸化能影响TrkA的激酶活性（3-6）。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（7-10）。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（8-13）。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（10）。

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.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-TrkA (Tyr490) Chemiluminescent Sandwich ELISA Kit #7024 detects endogenous levels of TrkA when phosphorylated at Tyr490 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.

Description

The PathScan® Phospho-TrkA (Tyr490) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of TrkA when phosphorylated at Tyr490 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 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, a Phospho-TrkA (Tyr490) Rabbit Detection Antibody is added to detect phospho-TrkA protein. Anti-rabbit IgG, HRP-linked Antibody 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 TrkA phosphorylated at Tyr490. 
 Antibodies in kit are custom formulations specific to kit.

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 (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).

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.

注意事项：

Low volume microplate * 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).