SignalSilence? CDK2 siRNA II

• 产品编号：CST-7417S      品牌：CST       原厂货号：7417S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

CDK2 siRNA II能够一直人类和小鼠CDK2蛋白表达。研究人员可以使用CST的SignalSilence® CDK2 siRNA II，采用RNA干扰（一种通过传递外源性的双链RNA分子到细胞中从而能选择性的沉默基因表达的方法）技术特异性的抑制CDK2 蛋白表达。CST所有的SignalSilence® siRNA产品都经过了严格的内部测试并采用Western分析证明其可以有效降低靶蛋白的表达。为了保证适当的结合效率，我们采用三苯甲基分析的方法逐个碱基的监测寡核苷酸的合成。随后通过亲和固相提取纯化寡核苷酸。退火的双链RNA进一步通过质谱分析来验证双链复合体的确切成分。每一批产品和前一批都要通过质谱分析进行比较以确定批次间最大限度的一致性。CST建议使用100 nM SignalSilence® CDK2 siRNA II转染48-72小时后裂解细胞。转染的具体步骤请参考转染试剂制造商提供的操作标准。使用过程中有任何问题请随时联系CST。周期蛋白依赖性激酶2(p33CDK2)是细胞周期机制的重要组成部分。类似于p34cdc2，激酶活性的调节经由磷酸化状态以及周期蛋白亚基和CDK抑制剂之间的关系。磷酸化抑制通常发生在14位和15位苏氨酸(1)。CDK2-周期蛋白复合体的抑制也与p27 Kip1和 p21 Waf1/Cip1之间的关系有关(2)。CDK2复合体的磷酸化需要cdc25磷酸酶和160位苏氨酸磷酸化引(由CDK7和cyclin H组成的CAK复合体介导)起的14位和15位苏氨酸去磷酸化。CDK2/周期蛋白E激酶活性对于G1到S期的转换和Rb蛋白的磷酸化具有重要意义。在S期，活化的CDK2/周期蛋白A复合体支配和磷酸化E2F，而活化的CDK2复合体持续在G2期的胞核中存在(5)。

1. Morgan, D.O. (1995) Nature 374, 131-4.
2. Poon, R.Y. et al. (1996) J Biol Chem 271, 13283-91.
3. Gu, Y. et al. (1992) EMBO J 11, 3995-4005.
4. Fesquet, D. et al. (1993) EMBO J 12, 3111-21.
5. Morgan, D.O. (1997) Annu Rev Cell Dev Biol 13, 261-91.

原厂资料：

Homology

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

Specificity / Sensitivity

CDK2 siRNA II will inhibit human and mouse CDK2 expression. 

Description

SignalSilence® CDK2 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit CDK2 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 CDK2 siRNA II 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

Cyclin-dependent kinase 2 (p33CDK2) is an important component of the cell cycle machinery. Like p34cdc2, kinase activity is regulated by phosphorylation state as well as association with a cyclin subunit and a CDK inhibitor. Inhibitory phosphorylation occurs on Thr14 and Tyr15 (1). Inhibition of CDK2-cyclin complexes can also be attributed to association with p27 Kip1 and p21 Waf1/Cip1 (2). Activation of CDK2 complexes requires dephosphorylation of Thr14 and Tyr15 by cdc25 phosphatase and phosphorylation of Thr160 (3), which is mediated by CAK, a complex of CDK7 and cyclin H (4). CDK2/cyclin E kinase activity is important for the G1 to S transition and phosphorylation of the Rb protein. During S-phase, active CDK2/cyclin A complexes predominate and phosphorylate E2F and the active CDK2 complex persists in the nucleus throughout G2 (5). 

1. Morgan, D.O. (1995) Nature 374, 131-4.
2. Poon, R.Y. et al. (1996) J Biol Chem 271, 13283-91.
3. Gu, Y. et al. (1992) EMBO J 11, 3995-4005.
4. Fesquet, D. et al. (1993) EMBO J 12, 3111-21.
5. Morgan, D.O. (1997) Annu Rev Cell Dev Biol 13, 261-91.

注意事项：

Storage: CDK2 siRNA II is supplied in RNAse-free water.
Aliquot and store at -20ºC.