Departments of Biophysics and Microbiology, UT Southwestern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2021;2263:161-181. doi: 10.1007/978-1-0716-1197-5_6.
Microscale thermophoresis (MST) has become a widely used technique to determine the K or EC of protein-ligand interactions. The method exploits the tendency of macromolecules to migrate along a thermal gradient (i.e., thermophoresis). Differences in thermophoresis as a function of the liganded state of a macromolecule can be measured and assembled into a binding curve that can be analyzed to yield K. In this protocol, we outline a simple experiment designed for new MST users, with the goal of using readily available, inexpensive materials to plan, execute, and analyze an MST experiment.
微尺度热泳（MST）已成为一种广泛使用的技术，用于确定蛋白质-配体相互作用的 K 或 EC。该方法利用大分子沿温度梯度（即热泳）迁移的趋势。可以测量大分子配体状态下热泳的差异，并将其组装成结合曲线，可用于分析得出 K 值。在本方案中，我们概述了一个专为新 MST 用户设计的简单实验，目的是使用现成的、廉价的材料来规划、执行和分析 MST 实验。
