Baker C S, Dunn M J, Yacoub M H
Department of Cardiothoraric Surgery, National Heart and Lung Institute, London, UK.
Electrophoresis. 1991 May;12(5):342-8. doi: 10.1002/elps.1150120505.
Protein purification and characterisation have been age-old problems for the biochemist. A new era has arisen with the advent of one- and two-dimensional gel electrophoresis and high sensitivity automated protein microsequencing. These two tools along with electroblotting have made it possible to separate and analyse complex protein mixtures. We studied six different membranes compatible with Edman degradation chemistry to determine their efficiencies at binding proteins electroblotted from one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Their overall blotting-sequencing properties were also evaluated. We found that the polyvinylidene difluoride-based membranes out-performed the glass-based and polypropylene-based membranes under our selected experimental conditions. The problems associated with electroblotting and microsequencing are discussed.
蛋白质的纯化与表征一直是生物化学家们长期面临的问题。随着一维及二维凝胶电泳和高灵敏度自动蛋白质微量测序技术的出现，一个新的时代已然来临。这两项技术与电转印技术一起，使得分离和分析复杂蛋白质混合物成为可能。我们研究了六种与埃德曼降解化学兼容的不同膜，以确定它们在结合从一维十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳电转印的蛋白质方面的效率。还评估了它们的整体印迹 - 测序特性。我们发现，在我们选定的实验条件下，基于聚偏二氟乙烯的膜比基于玻璃和聚丙烯的膜表现更优。文中还讨论了与电转印和微量测序相关的问题。
