Fischer Roman, Kessler Benedikt M
TDI Mass Spectrometry Laboratory, Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.
Proteomics. 2015 Apr;15(7):1224-9. doi: 10.1002/pmic.201400436. Epub 2015 Feb 5.
We describe a "gel-assisted" proteomic sample preparation method for MS analysis. Solubilized protein extracts or intact cells are copolymerized with acrylamide, facilitating denaturation, reduction, quantitative cysteine alkylation, and matrix formation. Gel-aided sample preparation has been optimized to be highly flexible, scalable, and to allow reproducible sample generation from 50 cells to milligrams of protein extracts. This methodology is fast, sensitive, easy-to-use on a wide range of sample types, and accessible to nonspecialists.
我们描述了一种用于质谱分析的“凝胶辅助”蛋白质组学样品制备方法。溶解的蛋白质提取物或完整细胞与丙烯酰胺共聚,促进变性、还原、半胱氨酸定量烷基化和基质形成。凝胶辅助样品制备已被优化,具有高度灵活性、可扩展性,并能从50个细胞到毫克级蛋白质提取物实现可重复的样品生成。该方法快速、灵敏、易于用于多种样品类型,且非专业人员也可操作。