低温电子显微
膜
膜蛋白
网格
计算机科学
化学
生物物理学
生物
生物化学
数学
几何学
作者
Magnus Bloch,Mònica Santiveri,Nicholas M. I. Taylor
标识
DOI:10.1007/978-1-0716-0373-4_16
摘要
Cryo-electron microscopy (cryo-EM) is a powerful tool for investigating the structure of macromolecules under near-native conditions. Especially in the context of membrane proteins, this technique has allowed researchers to obtain structural information at a previously unattainable level of detail. Specimen preparation remains the bottleneck of most cryo-EM research projects, with membrane proteins representing particularly challenging targets of investigation due to their universal requirement for detergents or other solubilizing agents. Here we describe preparation of negative staining and cryo-EM grids and downstream data collection of membrane proteins in detergent, by far the most common solubilization agent. This protocol outlines a quick and straightforward procedure for screening and determining the structure of a membrane protein of interest under biologically relevant conditions.
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