| 货号 | 6276S |
| 描述 | SignalSilence® Chk2 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Chk2 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.研究人员可以使用CST的SignalSilence® Chk2 siRNA I，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。Quality Control |
| 反应种属 | Human |
| 应用 | TFN |
| 供应商 | CST |
| 背景 | Chk2 is the mammalian orthologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases (1-3). The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68) each followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (4,5). After DNA damage by ionizing radiation (IR), UV irradiation or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR (5-7). The SQ/TQ cluster domain, therefore, seems to have a regulatory function. Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 on residues Thr383 and Thr387 in the activation loop of the kinase domain (8).Chk2是哺乳动物中和芽殖酵母Rad53和裂殖酵母Cds1检验点激酶同源的蛋白(1-3)。Chk2的氨基末端结构域包含有一个七丝氨酸或者苏氨酸残基(Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68)组成的系列，每个系列紧跟着谷氨酸(SQ 或TQ模体)。这些地方是已知的ATM/ATR激酶磷酸化的首选作用位点(4,5)。电离辐射(IR)引起DNA损伤之后，UV照射处理或者羟基脲处理，这个区域的68位苏氨酸和其他位点被ATM/ATR磷酸化(5-7)。因此，SQ/TQ簇结构域似乎具有调节功能。第68位苏氨酸的磷酸化是后续激活步骤的先决条件，激活归因于激酶结构域激活回路中Chk2第383位和387位苏氨酸残基的自体磷酸化(8)。 |
| 存放说明 | -20C |
| 参考文献 | Allen, J.B. et al. (1994) Genes Dev. 8, 2401-2415. Weinert, T.A. et al. (1994) Genes Dev. 8, 652-665. Murakami, H. and Okayama, H. (1995) Nature 374, 817-819. Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186. Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389-10394. Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765. Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934-5936. Lee, C.H. and Chung, J.H. (2001) J. Biol. Chem. 276, 30537-30541. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Chk2 siRNA I (+), using Chk2 (1C12) Mouse mAb #3440 and α-Tubulin (11H10) Rabbit mAb #2125. The Chk2 (1C12) Mouse mAb confirms silencing of Chk2 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.Western blot方法检测HeLa细胞提取物：100nM SignalSilence® Control siRNA (Unconjugated) #6568 (-)转染组，SignalSilence® Chk2 siRNA I (+)转染组，使用的抗体为Chk2 (1C12) Mouse mAb #3440 和α-Tubulin (11H10) Rabbit mAb #2125。 和对照抗体α-Tubulin (11H10) Rabbit mAb相比，Chk2 (1C12) Mouse mAb的检测结果证实 Chk2 的表达被有效沉默。 |
