Department of General Pediatrics, University Children's Hospital, Heinrich-Heine-University, Düsseldorf, Germany.
Anal Biochem. 2010 Dec 15;407(2):287-9. doi: 10.1016/j.ab.2010.08.009. Epub 2010 Aug 10.
Blue native polyacrylamide gel electrophoresis (BN-PAGE) is an essential tool for investigating mitochondrial respiratory chain complexes. However, with current BN-PAGE protocols for Caenorhabditis elegans (C. elegans), large worm amounts and high quantities of mitochondrial protein are required to yield clear results. Here, we present an efficient approach to isolate mitochondrial complex I (NADH:ubiquinone oxidoreductase) from C. elegans, grown on agar plates. We demonstrate that considerably lower amounts of mitochondrial protein are sufficient to isolate complex I and to display clear in-gel activity results. Moreover, we present the first complex I assembly profile for C. elegans, obtained by two-dimensional BN/SDS-PAGE.
蓝色 native 聚丙烯酰胺凝胶电泳 (BN-PAGE) 是研究线粒体呼吸链复合物的重要工具。然而，目前用于秀丽隐杆线虫 (C. elegans) 的 BN-PAGE 方案需要大量的线虫和高浓度的线粒体蛋白才能得到清晰的结果。在这里，我们介绍了一种从琼脂平板上生长的 C. elegans 中分离线粒体复合物 I (NADH:泛醌氧化还原酶) 的有效方法。我们证明，只需少量的线粒体蛋白即可分离复合物 I，并显示清晰的凝胶内活性结果。此外，我们通过二维 BN/SDS-PAGE 展示了 C. elegans 的第一个复合物 I 组装图谱。
Anal Biochem. 2010-8-10
J Chromatogr B Analyt Technol Biomed Life Sci. 2010-12-31
Zotero 插件