Borner Georg H H, Fielding Andrew B
Department of Proteomics and Signal Transduction, Max Plank Institute of Biochemistry, 82152 Martinsried, Germany;
Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, United Kingdom.
Cold Spring Harb Protoc. 2014 Nov 3;2014(11):1184-7. doi: 10.1101/pdb.prot083147.
HeLa cell lines can be experimentally manipulated using drugs or gene-silencing techniques such as RNA interference. Fractions enriched for clathrin-coated vesicles (CCVs) can be isolated from these cell lines and used to study the effects of these manipulations on the composition of CCVs. This protocol, originally developed in the laboratory of Margaret Robinson (Cambridge, United Kingdom), describes the preparation of a HeLa cell fraction that is enriched for a mixed population of CCVs and is suitable for analysis by mass spectroscopy, western blotting, or electron microscopy.
可以使用药物或基因沉默技术（如RNA干扰）对海拉细胞系进行实验操作。富含网格蛋白包被囊泡（CCV）的组分可从这些细胞系中分离出来，用于研究这些操作对CCV组成的影响。本实验方案最初由玛格丽特·罗宾逊实验室（英国剑桥）开发，描述了一种海拉细胞组分的制备方法，该组分富含混合的CCV群体，适用于质谱分析、蛋白质免疫印迹或电子显微镜分析。
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