| 货号 | 6241S |
| 描述 | SignalSilence® Chk1 siRNA from Cell Signaling Technology allows the researcher to specifically inhibit Chk1 expression using RNA interference, a method in which gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products are rigorously tested in-house and have been shown to reduce protein expression in specified cell lines.研究人员可以使用CST的SignalSilence® Chk1 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Chk1 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。 |
| 反应种属 | Human/Monkey |
| 应用 | TFN |
| 供应商 | CST |
| 背景 | Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 and occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for reentry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of Aurora B and BubR1 (8). Chk1 has emerged as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).Chk1激酶能够充当ATM/ATR激酶的下游并在DNA损伤检验点控制、胚胎发育和肿瘤抑制中发挥重要作用(1)。Chk1的激活涉及317位和345位丝氨酸的磷酸化，其发生响应DNA复制阻滞和某些类型的基因毒性应激反应(2)。尽管345位丝氨酸的磷酸化有助于伴随检验点激活的Chk1定位到核，317位丝氨酸的磷酸化连同位点特异性的PTEN磷酸化允许DNA复制停滞引起的细胞周期折返(4)。Chk1发挥其细胞周期检验点调节机制部分经由调控磷酸酶家族的cdc25。Chk1磷酸化cdc25A后靶向蛋白降解并通过14-3-3的结合抑制自身的活性(5)。激活的chk1能够通过216位丝氨酸的磷酸化失活cdc25C，阻塞cdc2的激活和有丝分裂转换的进入(6)。中心体的chk1被认为可以磷酸化cdc25B并抑制其激活CDK1-cyclin B1，藉此废除有丝分裂纺锤体的形成和染色质凝聚(7)。此外，Chk1通过调节Aurora B和BubR1在纺锤体检验点功能中发挥重要作用(8)。鉴于Chk1的抑制作用能够引起许多肿瘤细胞系的死亡，它已经成为肿瘤治疗中心的药物靶点(9)。 |
| 存放说明 | -20C |
| 计算分子量 | 54 |
| 参考文献 | Liu, Q. et al. (2000) Genes Dev 14, 1448-59. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39. Jiang, K. et al. (2003) J Biol Chem 278, 25207-17. Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16. Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97. Zeng, Y. et al. (1998) Nature 395, 507-10. Löffler, H. et al. (2006) Cell Cycle 5, 2543-7. Zachos, G. et al. (2007) Dev Cell 12, 247-60. Garber, K. (2005) J Natl Cancer Inst 97, 1026-8. Chen, Z. et al. (2003) Mol. Cancer Ther. 2(6) , 543-548. |
Western blot analysis of extracts from HeLa cells transfected with non-targeted (-) or targeted (+) siRNA. Chk1 was detected using the Chk1 Antibody #2345, and p42 was detected using the p42 MAPK Antibody #9108. The Chk1 Antibody confirms silencing of Chk1 expression, and the p42 MAPK Antibody was used to control for loading and specificity of Chk1 siRNA.Western blot方法检测转染非定向（-）或定向Chk1（+）siRNA的HeLa细胞。检测Chk1和p42的抗体分别为Chk1 Antibody #2345和p42 MAPK Antibody #9108 .Chk1 Antibody的检测结果证实 Chk1 的表达被有效沉默，p42 MAPK Antibody作为内对照和Chk1 siRNA作用特异性的对照。 | |
Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.荧光法检测转染后24小时活的HeLa细胞内的SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 。显示转染效率接近100%。 |
