SignalSilence? Fyn siRNA I

• 产品编号：CST-12473S      品牌：CST       原厂货号：12473S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® Fyn siRNA I抑制人源或猴源Fyn的表达。来自Cell Signaling Technology (CST)的SignalSilence^® Fyn siRNA I可以帮助研究者通过RNA干扰特异性地抑制Fyn的表达，这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence^® siRNA产品都是经过内部严格检测的，并且通过Western blot 分析证明确实能够减少目的蛋白的表达。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推荐使用100 nM SignalSilence^® Fyn siRNA I进行转染，48到72小时后对细胞进行裂解。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题，请随时联系CST。每小瓶可供100次转染，每次转染量相当于在转染24孔板时，每孔总体积为300μl培养基中siRNA的终浓度为100nM。Src酪氨酸激酶蛋白家族包括Src、Lyn、Fyn、Yes、Lck、Blk 和 Hck，对真核细胞的生长和分化过程发挥重要作用(1)。Src的活性是通过两个位点酪氨酸的磷酸化进行调节，但是具有相反的作用。在激酶活性环内的Tyr416 位点的磷酸化能够增加酶的活性，通过Csk磷酸化C-末端的Tyr527位点降低酶的活性(2)。Fyn 是一个分子量为59 kDa的Src家族酪氨酸激酶。Fyn的C-末端与Src具有很高的氨基酸序列相似度，但是在N-末端却有81个不同的氨基酸。Fyn蛋白合成后，其N-端在胞质多聚核糖体发生十八烷基化修饰，然后快速地转运至细胞质膜并发生棕榈酰化修饰(3)。与Src 蛋白中Tyr416和Tyr527周围氨基酸对应的氨基酸序列在Fyn 中是保守的，并因此可能也通过相似的磷酸化来调控。双重乙酰化修饰的Fyn 在细胞膜穴状内陷样的膜微区域聚集并与一系列的信号分子相互作用。Fyn的生物学功能多样，包括通过T细胞受体的信号传递, 调节脑的功能和粘连素介导的信号传递(4, 5)。在适当的靶组织中控制Fyn水平的变化可能促进相关疾病的治疗。

1. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
2. Hunter, T. (1987) Cell 49, 1-4.
3. Resh, M.D. (1998) Int J Biochem Cell Biol 30, 1159-62.
4. Nel, A.E. (2002) J Allergy Clin Immunol 109, 758-70.
5. Fukai, I. et al. (2000) Eur J Immunol 30, 3507-15.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® Fyn siRNA I inhibits human and monkey Fyn expression.

Description

SignalSilence® Fyn siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Fyn 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® Fyn 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

The Src family of protein tyrosine kinases, which includes Src, Lyn, Fyn, Yes, Lck, Blk, and Hck, are important in the regulation of growth and differentiation of eukaryotic cells (1). Src activity is regulated by tyrosine phosphorylation at two sites, but with opposing effects. While phosphorylation at Tyr416 in the activation loop of the kinase domain upregulates enzyme activity, phosphorylation at Tyr527 in the carboxy-terminal tail by Csk renders the enzyme less active (2).Fyn is a 59 kDa member of the Src family of tyrosine kinases. The carboxy terminus of Fyn shares extensive amino acid sequence homology with Src but is very different within the amino-terminal 81 amino acid residues. The Fyn protein is synthesized and N-myristoylated on cytosolic polysomes and then rapidly targeted to the plasma membrane, where it is palmitoylated (3). The corresponding sequences surrounding Tyr416 and Tyr527 of Src are conserved in Fyn and thus may be similarly regulated by phosphorylation. Dually acetylated Fyn clusters in caveolae-like membrane microdomains can interact with a variety of other signaling molecules. Fyn's biological functions are diverse and include signaling via the T cell receptor, regulation of brain function, and adhesion-mediated signaling (4, 5). Alteration of the levels of Fyn in appropriate target tissues may lead to better treatments for some related diseases.

1. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
2. Hunter, T. (1987) Cell 49, 1-4.
3. Resh, M.D. (1998) Int J Biochem Cell Biol 30, 1159-62.
4. Nel, A.E. (2002) J Allergy Clin Immunol 109, 758-70.
5. Fukai, I. et al. (2000) Eur J Immunol 30, 3507-15.

注意事项：

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