Substitution of Deoxycholate with the Amphiphilic Polymer Amphipol A8-35 Improves the Stability of Large Protein Complexes during Native Electrophoresis

Abstract Native polyacrylamide gel electrophoresis (PAGE) is a powerful technique for protein complex separation that retains both their activity and structure. In photosynthetic research, native-PAGE particularly useful given complexes are generally large in size, ranging from 200 kD to 1 MD or more. Recently, it has been reported the addition of amphipol A8-35 solubilized samples improved stability. previous study, we found could substitute sodium deoxycholate (DOC), conventional electrophoretic carrier, clear-native (CN)-PAGE. this present optimization amphipol-based CN-PAGE. We ratio ?-dodecyl maltoside, detergent commonly used solubilize complexes, was critical resolving machinery addition, LHCII dissociation PSII–LHCII effectively prevented by CN-PAGE compared with DOC-based Our data strongly suggest majority vivo forms C2S2M2 at least Arabidopsis Physcomitrella. The other might appear owing PSII during sample preparation electrophoresis, which be after solubilization thylakoid membranes. These results may better alternative study labile complexes.