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Ultra-filtration (to wash out un-reacted free trisulfides) Trisulfide-pretreated HGF solutions were loaded into Amicon ultra-filtration devices (with a 10-kDa MWCO polyethersulfone (PES)-membrane) and spun at the recommended centrifugal force (typically 6000-7000 rpm) for 10–30 min at 4℃ or until the desired volume was reached