The pellet was resuspended in 10
ml of resuspension buffer (10 mM Mes-Tris (pH 7.5),
250 mM sucrose, 5% glycerol (v/v), 1 mM EDTA, 1 mM
DTT, 0.1 mM PMSF),layered on a 25/50% sucrose step
gradient made up in gradient buffer (5 mM Mes-Tris
(pH 7.5), 1 mM EDTA, 1 mM DTT and 0.1 mM
PMSF).
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