cells were washed, resuspended in ice-cold PBS, and disrupted with a tissue homogenizer (three 10 second periods) in the presence of anti-protease cocktail (Sigma Aldrich). Whole cells, nuclei, and other large debris were cleared by centrifugation at 300 × g for 10 min at 4°C and the particulate fraction (crude membrane fraction) of the cleared supernatant was separated by microcentrifugation at maximum speed for 30 min. The protein content was determined by standard BSA protein assay.不知道是否适合你的实验

方法很多，可以用去垢剂分布抽提，见蛋白质与蛋白质组学实验指南，也可以用超速离心的方法分离，条件是您的离心机可达到100000g，关于该方法文献上比比皆是。最简单的方法，pierce卖试剂盒，我用过效果很好，不过很贵

有专门的提取试剂盒，建议用进口的