千赫兹帧率下的高分辨率单分子驱动蛋白分析
High-Resolution Single-Molecule Kinesin Assays at kHz Frame Rates.
作者信息
Mickolajczyk Keith J, Hancock William O
机构信息
Biomedical Engineering Department, Pennsylvania State University, University Park, PA, USA.
出版信息
Methods Mol Biol. 2018;1805:123-138. doi: 10.1007/978-1-4939-8556-2_7.
Abstract
This chapter describes methods for high-speed, unloaded, in vitro single-molecule kinesin tracking experiments. Instructions are presented for constructing a total internal reflection dark-field microscope (TIRDFM) and labeling motors with gold nanoparticles. An AMP-PNP unlocking assay is introduced as a specialized means of capturing processive events in a reduced field of view. Finally, step-finding tools for analyzing high frame-rate tracking data are described.
摘要
本章介绍了用于高速、无负载的体外单分子驱动蛋白追踪实验的方法。文中给出了构建全内反射暗场显微镜（TIRDFM）以及用金纳米颗粒标记马达蛋白的说明。引入了一种AMP-PNP解锁测定法，作为在缩小视野中捕捉持续运动事件的一种特殊手段。最后，描述了用于分析高帧率追踪数据的步长查找工具。
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