Preparation of highly purified F-actin-depolymerizing factor of human serum.
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% (NH4)2SO4 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.
Thorstensson
R
R
Sterky
C
C
Norberg
R
R
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000097:blood*; D001798:Blood Proteins; D002135:Calcium-Binding Proteins / Q000302:isolation & purification; D002352:Carrier Proteins / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D051345:Destrin; D004591:Electrophoresis, Polyacrylamide Gel; D018260:Gelsolin; D006801:Humans; D007525:Isoelectric Focusing; D008840:Microfilament Proteins / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*
D051339:Actin Depolymerizing Factors; D000199:Actins; D001798:Blood Proteins; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; C495824:DSTN protein, human; D051345:Destrin; D018260:Gelsolin; D008840:Microfilament Proteins; D011506:Proteins; C016953:brevin