Detection of S-phase cell cycle progression using 5-ethynyl-2′-deoxyuridine incorporation with click chemistry an alternative to using 5-bromo-2′-deoxyuridine antibodies
Detection of S-phase cell cycle progression using 5-ethynyl-2′-deoxyuridine incorporation with click chemistry an alternative to using 5-bromo-2′-deoxyuridine antibodies
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DOI：
10.2144/000112812
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发表时间：
2008-06-01
期刊：
影响因子：
2.7
通讯作者：
Salic, Adrian
中科院分区：
文献类型：
--
作者：
Buck, Suzanne B.;Bradford, Jolene;Salic, Adrian
细胞的 5-bromo-2'-deoxvuridine (BrdU) 标记和抗体染色已成为直接测量 S 期细胞的标准方法。所描述的是一个。基于点击化学应用的改进的 S 期细胞周期进展检测方法，点击化学是末端炔烃和叠氮化物之间 Huisgen [3+2] 环加成的铜 (1) 催化变体。 5-乙炔基-2'-脱氧尿苷 (EdU) 是胸苷的核苷类似物，与 BrdU 一样，在活跃的 DNA 合成过程中掺入 DNA。虽然 BrdU 测定需要对螺旋 DNA 结构进行严格的化学或酶促破坏，以便通过抗 BrdU 抗体直接测量 S 期细胞，但 EdU 方法则不需要。消除这一要求可以保留螺旋 DNA 结构和其他细胞表面表位、减少测定时间并提高重现性。
The 5-bromo-2'-deoxvuridine (BrdU) labeling of cells followed by antibody staining has been the standard method for direct measurement of cells in the S-phase. Described is an. improved method for the detection of S-phase cell cycle progression based upon the application of click chemistry, the copper(1)-catalyzed variant of the Huisgen [3+2] cycloaddition between a terminal alkyne and an azide. 5-ethynyl-2'-deoxyuridine (EdU) is a nucleoside analog of thymidine that is incorporated into DNA during active DNA synthesis, just like BrdU. While the BrdU assay requires harsh chemical or enzymatic disruption of helical DNA structure to allow for direct measurement of cells in the S-phase by the anti-BrdU antibody, the EdU method does not. Elimination of this requirement results in the preservation of helical DNA structure and other cell surface epitopes, decreased assay time, and increased reproducibility.