SignalSilence? TrkA siRNA I

• 产品编号：CST-6613S      品牌：CST       原厂货号：6613S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司生产的SignalSilence®TrkA siRNA I使得研究者可以使用RNA干扰技术特异性的抑制TrkA的表达。这种技术通过将双链RNA分子送入细胞而选择性的沉默某个基因的表达。CST所有SignalSilence®siRNA产品已经在内部经过严格测试，经western分析确能减少目标蛋白的表达。通过三苯分析对寡核苷酸合成进行逐个碱基的监测，确保了正确的耦联效率。然后用亲和固相萃取纯化寡核苷酸。退火的RNA进一步通过质谱分析，以验证其确切成分。每批都将和以前批次进行质谱比对，以确保批次间最大的一致性。CST公司推荐您在细胞裂解前用100nM TrkA siRNA I进行48-72小时的转染。转染的流程请遵循转染试剂制造商提供的操作流程。使用中如有任何问题，请与CST联系。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.

原厂资料：

Description

SignalSilence® TrkA siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit TrkA expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM TrkA siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

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.

注意事项：

Storage: TrkA siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.