SignalSilence? cdc2 siRNA II

• 产品编号：CST-3600S      品牌：CST       原厂货号：3600S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

研究人员可以使用CST的SignalSilence® cdc2 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cdc2 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® cdc2 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。 cdc2蛋白激酶的激活参与调节真核细胞进入有丝分裂，cdc2的激活受多个步骤的调控，包括周期蛋白的结合、cdc2苏氨酸161位的磷酸化(1)。然而，在细胞进入有丝分裂的过程中，激活cdc2的决定性步骤是苏氨酸14位和15位的去磷酸化(2)。Wee1 和 Myt1蛋白激酶通过磷酸化cdc2苏氨酸14位和15位，抑制cdc2活性(3,4)。而cdc25磷酸酯酶可能起到使cdc2苏氨酸14位和15位去磷酸化从而激活cdc2的作用(1,5)。

1. Atherton-Fessler, S. et al. (1994) Mol Biol Cell 5, 989-1001.
2. Norbury, C. et al. (1991) EMBO J 10, 3321-9.
3. McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
4. Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.
5. Hunter, T. (1995) Cell 80, 225-36.

原厂资料：

Description

SignalSilence® cdc2 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit cdc2 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® cdc2 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

The entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Thr14 and Tyr15 (2). Phosphorylation at Thr14 and Tyr15, resulting in inhibition of cdc2, can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).

1. Atherton-Fessler, S. et al. (1994) Mol Biol Cell 5, 989-1001.
2. Norbury, C. et al. (1991) EMBO J 10, 3321-9.
3. McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
4. Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.
5. Hunter, T. (1995) Cell 80, 225-36.

注意事项：

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