【摘要】 目的:分析最新近红外荧光标记(NIRF)电泳迁移率变动分析(EMSA)技术的优缺点,在此基础上对NIRF-EMSA技术进行实验方法的改进。方法:设计了3种NIRF-EMSA实验方法,即扫胶法、染胶法和扫膜法,通过实验比较了3种方法的实验程序复杂性、优缺点及检测成本,并将3种方法与化学发光EMSA进行了比较。结果:采用3种方法都可以取得良好的EMSA检测效果,但比较而言,间接扫膜法不仅效果良好,而且实验成本最低。结论:间接扫膜法是值得推广应用的最经济便捷的基于Odyssey红外荧光成像系统的近红外荧光EMSA实验方法。更多还原

【Abstract】 Objective:The electrophoresis mobility shift assay(EMSA) is golden standard technique for studying the interaction of DNA and protein.The merits and limitations of the latest near-infrared fluorescence-labeled EMSA(NIRF-EMSA) were briefly analyzed.On this base,experiments were designed to improve the NIRF-EMSA protocol for routine application.Methods:We tried three NIRF-EMSA protocols,including scanning gel,staining gel and scanning membrane,and systematically evaluated the complexity of experimental procedures,merits and disadvantages,and the assay cost.At the same time,three NIRF-EMSA approaches were compared with chemiluminescent EMSA.Results:Three protocols all allowed for better detection results.However,in comparison,scanning membrane was the most cost-effective approach.Conclusion:Scanning membrane is the most facile and cost-effective NIRF-EMSA method worthy to be applied to routine experiment in the future.更多还原