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For rotary shadowing, sample proteins were suspended in
solutions of 50% glycerol containing 0.4 M ammonium acetate,
pH 7.2. Sample suspensions were sprayed on mica sheets and then
the platinum¨Ccarbon was shadowed, while rotating with an electron
beam evaporator (JFD-9010, JOEL, Tokyo, Japan). Following
these treatments, the replica was floated off on water and picked
up on a copper grid. Samples were observed under a transmission
electron microscope.
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