Membrane protein isolation and structure determination in cell-derived membrane vesicles | Proceedings of the National Academy of Sciences thumbnail
Membrane protein isolation and structure determination in cell-derived membrane vesicles | Proceedings of the National Academy of Sciences
www.pnas.org
The bound vesicles were eluted by incubating with HRV-3C protease at a final concentration of ~0.014 mg/mL at 4 °C for 2 h. Dounce homogenizer for ~30 to 60 strokes We added EDTA to potentially reduce divalent ion–mediated vesicle and protein aggregation, but we have not tested whether incorporating
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  • The bound vesicles were eluted by incubating with HRV-3C protease at a final concentration of ~0.014 mg/mL at 4 °C for 2 h.
  • Dounce homogenizer for ~30 to 60 strokes
  • We added EDTA to potentially reduce divalent ion–mediated vesicle and protein aggregation, but we have not tested whether incorporating EDTA is necessary.
  • 1 mM fluorinated fos-choline-8
  • GFP nanobody–conjugated affinity resin

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