Astbury Centre for Structural Molecular Biology, School of Biomedical Science, University of Leeds, Leeds, UK.
Methods Mol Biol. 2020;2168:263-269. doi: 10.1007/978-1-0716-0724-4_12.
Size-exclusion chromatography coupled to multiangle laser light scattering (SEC-MALLS) is the perfect method to determine the oligomeric state of membrane proteins as this method works in solution and is totally independent from prior assumptions such as detergent-to-protein ratio or the shape of the protein. In a relatively short time (ca. 30 min), the molecular mass and quality of a membrane protein preparation can be determined. Here, I provide a detailed protocol on how to perform a SEC-MALLS run and show exemplary chromatograms and their analysis.
尺寸排阻色谱法与多角度激光光散射（SEC-MALLS）相结合是确定膜蛋白寡聚状态的完美方法，因为该方法在溶液中进行，完全独立于诸如去污剂与蛋白质的比例或蛋白质形状等先前的假设。在相对较短的时间（约 30 分钟）内，可以确定膜蛋白制剂的分子量和质量。在这里，我提供了一个详细的协议，说明如何进行 SEC-MALLS 运行，并展示了示例色谱图及其分析。
