Selected article for: "electron microscope and uranyl acetate"
Author: Wilton T. Snead; Wade F. Zeno; Grace Kago; Ryan W. Perkins; J Blair Richter; Chi Zhao; Eileen M. Lafer; Jeanne C. Stachowiak
Title: BAR scaffolds drive membrane fission by crowding disordered domains
  • Document date: 2018_3_4
    • ID: drqseaaa_53
    Snippet: Vesicles for electron microscopy were composed of 5 mol% PtdIns(4,5)P 2 , 15 mol% DOPS, and 80 mol% DOPC. Dried lipid films were hydrated in 20 mM MOPS pH 7.35, 150 mM NaCl, 0.5 mM EGTA and EDTA buffer and extruded though a 200 nm pore filter (Whatman). Proteins were diluted to the indicated concentrations in the same MOPS buffer with 5 mM TCEP and incubated with vesicles at 37 °C for 30 min (Amph-FL and N-BAR) or 60 min (FCHo1-FL and F-BAR). Th.....
    Document: Vesicles for electron microscopy were composed of 5 mol% PtdIns(4,5)P 2 , 15 mol% DOPS, and 80 mol% DOPC. Dried lipid films were hydrated in 20 mM MOPS pH 7.35, 150 mM NaCl, 0.5 mM EGTA and EDTA buffer and extruded though a 200 nm pore filter (Whatman). Proteins were diluted to the indicated concentrations in the same MOPS buffer with 5 mM TCEP and incubated with vesicles at 37 °C for 30 min (Amph-FL and N-BAR) or 60 min (FCHo1-FL and F-BAR). The vesicle concentration was 1 mM in experiments with Amph-FL, FCHo1-FL, and F-BAR, and 0.1 mM in experiments with N-BAR and in protein-free controls. 5 µL of the mixture was placed onto a glow-discharged, 300 square mesh, carbon-coated grid and stained with 2% uranyl acetate (Electron Microscopy Sciences; Hatfield, PA, USA). Images were collected on a Tecnai Spirit BioTwin T12 electron microscope (Tecnai; Hillsboro, OR, USA). Vesicle and tubule diameters were measured using ImageJ software. All rights reserved. No reuse allowed without permission.

    Search related documents: