Open AccessPreparation of highly purified F‐actin‐depolymerizing factor of human serum
Author(s) -
THORSTENSSON Rigmor,
STERKY Catharina,
NORBERG Renée
Publication year - 1985
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.0014-2956.1985.00637.x
Subject(s) - chromatofocusing , proteolysis , affinity chromatography , chromatography , ion chromatography , specific activity , yield (engineering) , chemistry , sepharose , ion exchange , reproducibility , biochemistry , size exclusion chromatography , enzyme , ion , organic chemistry , materials science , metallurgy
F‐actin depolymerizing factor (ADF) of human serum was purified 250–400‐fold to more than 98% purity with high reproducibility. The purification included (1) 30–50% (NH 4 ) 2 SO 4 precipitation, (2) ion‐exchange chromatography on DEAE‐Sepharose, (3) chromatofocusing on Polybuffer exchanger 94 and (4) affinity chromatography on ConA‐Sepharose. The recovery of ADF was estimated to be 20–30% whereas the ADF activity yield was 5–17%. The lower activity yield was thought to be due‐partly to proteolysis and partly to destabilization of highly purified ADF.