SignalSilence? Cyclin D1 siRNA I

• 产品编号：CST-12962S      品牌：CST       原厂货号：12962S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® Cyclin D1 siRNA I可以抑制人和猴的cyclin D1表达。Cell Signaling Technology (CST) 公司的SignalSilence® Cyclin D1 siRNA I 帮助研究人员使用RNA干扰技术特异性抑制cyclin D1表达，该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试，western分析显示能够能够降低目标蛋白的表达。寡核苷酸合成经过了三苯甲基的逐碱基分析以确保耦合效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。CST建议转染100 nM SignalSilence® Cyclin D1 siRNA I，48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题，可以随时咨询CST。 每管试剂能进行100次转染，该转染次数以总体积为300μl 的1个24孔板孔中siRNA终浓度100 nM来进行计算。细胞周期依赖性激酶CDK4和CDK6的活性受到T-loop磷酸化，它们的细胞周期蛋白搭档（D-type cyclins）的丰度，以及与Cip/Kip 或INK家族蛋白的CDK抑制子的结合调控（1）。失活的三聚体cyclin D/CDK4 和p27 Kip1需要细胞外有丝分裂原激活以释放和降解p27，伴随细胞周期打那笔D水平升高以影响通过限制点和Rb-依赖的S-期进入（2）。cyclin D/CDK4的活性复合物靶向成视网膜细胞瘤蛋白磷酸化，允许E2F转录因子释放激活G1/S-期基因表达（3）.生长因子水平撤退会下降周期蛋白D的水平是通过下调蛋白表达水平和磷酸化依赖的的蛋白降解来继续拧的（4）。

1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
2. Sherr, C.J. (1996) Science 274, 1672-1677.
3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence^® Cyclin D1 siRNA I inhibits human and monkey cyclin D1 expression.

Description

SignalSilence^® Cyclin D1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit cyclin D1 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^® Cyclin D1 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

Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to affect progression through the restriction point and Rb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of protein expression and phosphorylation-dependent degradation (4).

1. Hirai, H. et al. (1995) Mol. Cell. Biol. 15, 2672-2681.
2. Sherr, C.J. (1996) Science 274, 1672-1677.
3. Lukas, J. et al. (1996) Mol. Cell. Biol. 16, 6917-6925.
4. Diehl, J.A. et al. (1997) Genes Dev. 11, 957-972.

注意事项：

Storage: Cyclin D1 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.