Manadas Bruno J, Vougas Konstantinos, Fountoulakis Michael, Duarte Carlos B
Department of Zoology, Center for Neuroscience and Cell Biology, Dept. of Zoology, University of Coimbra, Coimbra, Portugal.
Electrophoresis. 2006 May;27(9):1825-31. doi: 10.1002/elps.200500757.
Protein precipitation with TCA followed by acetone washing is frequently used to clean samples before 2-DE. However, the difficulty in solubilizing TCA-precipitated proteins causes some variability in 2-D gels and makes it difficult to detect some proteins. In this work we show that sonication of the samples, after TCA precipitation followed by elution in sample buffer, increases total protein recovery, and improves reproducibility and matching ratios between gels when analyzed by specialized software.
在进行二维电泳(2-DE)之前,常用三氯乙酸(TCA)沉淀蛋白质,随后用丙酮洗涤来清理样品。然而,TCA沉淀的蛋白质难以溶解,这会导致二维凝胶出现一些差异,并且难以检测到某些蛋白质。在本研究中,我们表明,在TCA沉淀样品后,于样品缓冲液中洗脱,再对样品进行超声处理,可提高总蛋白回收率,并在使用专业软件分析时提高凝胶之间的重现性和匹配率。