Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP 13083-862, Brazil.
Anal Biochem. 2011 Nov 1;418(1):158-60. doi: 10.1016/j.ab.2011.06.044. Epub 2011 Jul 13.
To isolate mitochondrial complexes, we have combined elements from the classic Laemmli protocol and blue native polyacrylamide gel electrophoresis (BN-PAGE) methods to develop a straightforward modified native electrophoresis protocol. This modified protocol presented good resolution for native electrophoresis of inner mitochondrial membrane proteins, where bands were easily visualized with no leftover stain or gel lanes overlap. Enzymatic tests revealed that complexes I and V remain active in the gel. This protocol, designed to overcome specific limitations of the standard protocols, provides a potential methodology to study membrane proteins in their functional form.
为了分离线粒体复合物，我们结合了经典的 Laemmli 方案和蓝色 native 聚丙烯酰胺凝胶电泳（BN-PAGE）方法中的元素，开发了一种简单的改良天然电泳方案。该改良方案对内质网膜蛋白的天然电泳具有良好的分辨率，无需多余的染色或凝胶条带重叠，即可轻松观察到条带。酶学测试表明复合物 I 和复合物 V 在凝胶中仍然保持活性。该方案旨在克服标准方案的特定限制，为研究功能形式的膜蛋白提供了一种潜在的方法。
