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10.1194/jlr.R500013-JLR200 51

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For experiments with IMVs and for the purification of EtfD, the membrane fraction was resuspended in 15 mL (assays) or 40 mL (protein purification) S-buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 20% (v/v) glycerol, 5 mM MgSO 4 , 5 mM benzamidine hydrochloride, 5 mM aminocaproic acid, 1 mM PMSF)

Following overnight incubation with the primary antibodies, membranes were washed five times for 15 min in TBST with mild shaking