Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, USA.
Electrophoresis. 2010 Jun;31(12):1934-6. doi: 10.1002/elps.201000019.
Extracting and concentrating mitochondrial protein complexes from gel strips after blue native PAGE (BN-PAGE) can be daunting tasks using the traditional methods, such as electroelution, passive diffusion and centrifugal concentration. We present a simplified gel electrophoresis method to concentrate mitochondrial protein complexes with excellent recovery rate. Mitochondrial complex I present in a long gel strip from BN-PAGE can be easily concentrated into a 0.8 cm gel strip when a second BN-PAGE is performed with a Y-shaped gel and the addition of 0.01% n-dodecyl beta-D-maltoside and 0.001% SDS in the cathode buffer. Once completed, the concentrated protein complex in the gel strip is ready for SDS-PAGE or proteomic studies.
从蓝色 nativePAGE（BN-PAGE）凝胶条中提取和浓缩线粒体蛋白复合物，使用传统方法（如电洗脱、被动扩散和离心浓缩）可能会很困难。我们提出了一种简化的凝胶电泳方法，可实现线粒体蛋白复合物的浓缩，具有出色的回收率。当在阴极缓冲液中添加 0.01%n-十二烷基-β-D-麦芽糖苷和 0.001%SDS 并使用 Y 形凝胶进行第二次 BN-PAGE 时，来自 BN-PAGE 的长凝胶条中的线粒体复合物 I 可轻松浓缩到 0.8cm 的凝胶条中。完成后，凝胶条中的浓缩蛋白复合物即可用于 SDS-PAGE 或蛋白质组学研究。
