Max Planck Institute of Biochemistry, Department of Molecular Structural Biology, Martinsried, Germany.
Nat Methods. 2012 Jan 8;9(2):182-4. doi: 10.1038/nmeth.1840.
We report a simple and generic method for the direct transfer of protein complexes separated by native gel electrophoresis to electron microscopy grids. After transfer, sufficient material remains in the gel for identification and characterization by mass spectrometry. The method should facilitate higher-throughput single-particle analysis by substantially reducing the time needed for protein purification, as demonstrated for three complexes from Thermoplasma acidophilum.
我们报告了一种简单通用的方法,可将通过天然胶电泳分离的蛋白质复合物直接转移到电子显微镜网格上。转移后,凝胶中仍留有足够的材料,可通过质谱进行鉴定和表征。该方法应该通过大大减少蛋白质纯化所需的时间,从而促进高通量的单颗粒分析,我们已经用嗜酸性热原体的三个复合物证明了这一点。