SignalSilence? FGF Receptor 2 siRNA I

• 产品编号：CST-12600S      品牌：CST       原厂货号：12600S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® FGF Receptor 2 siRNA I抑制人和猴子FGF受体2的表达。CST SignalSilence® FGF Receptor 2 siRNA I允许研究者使用RNA干扰技术特定地抑制FGF受体2表达，使用这种方法可以将双链RNA分子转至细胞内，选择性沉默基因的表达。所有CST SignalSilence® siRNA产品都经过严格的内部测试，并用western方法检测证明可以降低靶蛋白的表达。通过三苯甲基分析每个碱基以监测寡核苷酸的合成，确保合适的配对效率。随后寡核苷酸通过亲和固相萃取法纯化。退火的RNA双链通过质谱分析来证实其精确的组成。每一批产品都通过质谱分析与前面的产品进行比较，来保证不同批次之间的最大一致性。CST建议在细胞裂解前48-72h，用100 NM SignalSilence® FGF Receptor 2 siRNA I进行转染。转染程序参照转染试剂制造商所提供的操作指南。在使用上有任何问题，请随时联系CST。每管含有相当于100个转染的量，这相当于每个24孔板，每次转染用100 nM的终浓度siRNA，每孔总体积为300μl。成纤维细胞生长因子 (FGFs)通过细胞表面的酪氨酸激酶受体传递信号，对靶细胞产生促有丝分裂和血管生成的效应。已知FGF受体家族有四个成员：FGFR-1 (flg), FGFR-2 (bek, KGFR), FGFR-3和FGFR-4。每个受体都有一个胞外配体结合区、跨膜区和一个胞内激酶区（1）。在结合配体和二聚化之后，受体的特定的酪氨酸位点发生磷酸化（2）。在FGFR-1的羧基端有7个酪氨酸位点能被磷酸化：Tyr463, 583, 585, 653, 654, 730和 766。Tyr653 和Tyr654对激活的FGFR的催化活性十分重要，对信号传导也是必要的(3)。其它的磷酸化酪氨酸位点能够为下游信号分子，如Crk 和PLCγ，提供结合位点(4,5)。FGFR2有一些剪接异构体，配体特异性主要是由选择性剪接的外显子8（IIIb）和9（IIIc）决定的。选择性剪接是细胞类型特异性的，从而导致亚型在各种组织中的分布和生物活性（6,7）。调查研究表明，在相应的FGFR2基因突变导致特点是面部及肢体的缺陷的综合征，包括拉德综合征，克鲁综合症，BEARE史蒂文森表皮Gyrata综合征，菲佛综合症，亚伯综合症和杰克逊 - 魏斯综合征（8-10）。调查还发现在胃，子宫内膜癌和乳腺癌中，FGFR2的突变和表达改变（11）。

1. Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
2. Reilly, J.F. et al. (2000) J Biol Chem 275, 7771-8.
3. Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89.
4. Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78.
5. Larsson, H. et al. (1999) J Biol Chem 274, 25726-34.
6. Muh, S.J. et al. (2002) J Biol Chem 277, 50143-54.
7. Coutts, J.C. and Gallagher, J.T. (1995) Immunol Cell Biol 73, 584-9.
8. Jeftha, A. et al. (2004) J Clin Pediatr Dent 28, 173-6.
9. Wilkinson, C.C. et al. (2012) Childs Nerv Syst 28, 1221-6.
10. Slavotinek, A. et al. (2009) Am J Med Genet A 149A, 1814-7.
11. Katoh, M. (2009) J Invest Dermatol 129, 1861-7.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Monkey

Specificity / Sensitivity

SignalSilence® FGF Receptor 2 siRNA I inhibits human and monkey FGF Receptor 2 expression.

Description

SignalSilence® FGF Receptor 2 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit FGF Receptor 2 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 SignalSilence® FGF Receptor 2 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. 
 Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

Fibroblast growth factors (FGFs) produce mitogenic and angiogenic effects in target cells by signaling through cell surface receptor tyrosine kinases. There are four members of the FGF receptor family: FGFR1 (flg), FGFR2 (bek, KGFR), FGFR3, and FGFR4. Each receptor contains an extracellular ligand binding domain, a transmembrane domain, and a cytoplasmic kinase domain (1). Following ligand binding and dimerization, the receptors are phosphorylated at specific tyrosine residues (2). Seven tyrosine residues in the cytoplasmic tail of FGFR1 can be phosphorylated: Tyr463, 583, 585, 653, 654, 730, and 766. Tyr653 and Tyr654 are important for catalytic activity of activated FGFR and are essential for signaling (3). The other phosphorylated tyrosine residues may provide docking sites for downstream signaling components such as Crk and PLCγ (4,5).FGFR2 has several splicing isoforms with ligand specificity largely determined by alternative splicing of exons 8 (IIIb) and 9 (IIIc). Alternative splicing is cell type specific, resulting in isoforms showing various tissue distribution and biological activities (6,7). Research studies have shown that mutations in the corresponding FGFR2 gene cause syndromes characterized by facial and limb defects, including LADD Syndrome, Crouzon Syndrome, Beare-Stevenson Cutis Gyrata Syndrome, Pfeiffer Syndrome, Apert Syndrome, and Jackson-Weiss Syndrome (8-10). Investigators have also observed mutations and altered expression of FGFR2 in cases of gastric, endometrial, and breast cancer (11).

1. Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
2. Reilly, J.F. et al. (2000) J Biol Chem 275, 7771-8.
3. Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89.
4. Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78.
5. Larsson, H. et al. (1999) J Biol Chem 274, 25726-34.
6. Muh, S.J. et al. (2002) J Biol Chem 277, 50143-54.
7. Coutts, J.C. and Gallagher, J.T. (1995) Immunol Cell Biol 73, 584-9.
8. Jeftha, A. et al. (2004) J Clin Pediatr Dent 28, 173-6.
9. Wilkinson, C.C. et al. (2012) Childs Nerv Syst 28, 1221-6.
10. Slavotinek, A. et al. (2009) Am J Med Genet A 149A, 1814-7.
11. Katoh, M. (2009) J Invest Dermatol 129, 1861-7.

注意事项：

Storage: FGF Receptor 2 siRNA I is supplied in RNAse-free
water. Aliquot and store at -20ºC.