Levental K R, Levental I
Department of Integrative Biology and Pharmacology, University of Texas Health Science Center at Houston Medical School, 6431 Fannin Street, Houston, TX, 77030, USA,
Methods Mol Biol. 2015;1232:65-77. doi: 10.1007/978-1-4939-1752-5_6.
Although investigation into the structure of eukaryotic cell membranes has been an intense focus of cell biology for the past two decades, definitive insights have been limited by the lack of coherent methods for the isolation of specific organelle membranes and the identification of membrane subdomains. Here we describe a method for the isolation of mammalian cell plasma membranes as Giant Plasma Membrane Vesicles (GPMVs) and strategies for imaging membrane lateral structure and quantification of protein partitioning between coexisting domains by fluorescence microscopy.
尽管在过去二十年中，对真核细胞膜结构的研究一直是细胞生物学的一个重点，但由于缺乏用于分离特定细胞器膜和鉴定膜亚结构域的连贯方法，明确的见解受到了限制。在这里，我们描述了一种将哺乳动物细胞质膜分离为巨型质膜囊泡（GPMV）的方法，以及通过荧光显微镜对膜侧向结构进行成像和定量共存结构域之间蛋白质分配的策略。
