Sequential treatment by phosphotungstic acid and uranyl acetate enhances the adherence of lipid membranes and membrane proteins to hydrophobic EM grids.
A simple procedure for screening by electron microscopic observations of conditions for the reconstitution of membrane proteins into lipid bilayers is described. This procedure consists of a 5-10 s treatment of electron microscopic grids, to which the sample has already been applied, with 1% phosphotungstic acid before proceeding with final staining in uranyl acetate. The method substantially enhances the adherence of lipid membranes and membrane protein particles to hydrophobic collodion/carbon grids.
Biochemistry/Biophysics Department, Washington State University, Pullman 99164-4660.
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Barnakov
D000268:Adhesiveness; D008051:Lipid Bilayers / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron / Q000379:methods*; D009942:Organometallic Compounds; D010772:Phosphotungstic Acid
D008051:Lipid Bilayers; D008565:Membrane Proteins; D009942:Organometallic Compounds; D010772:Phosphotungstic Acid; C005460:uranyl acetate