Rapid, activity-guided isolation of sex-limited protein (Slp) from mouse serum by fractionated precipitation and high-performance liquid chromatography.
We have recently demonstrated that sex-limited protein (Slp) plays a key role in an EDTA-resistant mouse complement activation pathway. A rapid procedure, utilizing classical chromatography methods on an FPLC system, was developed for the isolation of functionally active Slp. The method is based on the fractionated precipitation of serum by polyethylene glycol 6000, followed by heparin Sepharose Cl-6B affinity chromatography, Mono Q anion exchange and Superose 12 gel filtration. The isolation of Slp was monitored by a hemolytic assay. The procedure resulted in the purification of Slp, which by SDS-PAGE gave a single band of M(r) 2000,000 under non-reducing conditions, and under reducing conditions three bands corresponding to M(rs) of 105,000, 76,000 and 37,000.
Van den Berg
C W
CW
Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, UK.
Van Dijk
H
H
D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification*; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003181:Complement C4 / Q000302:isolation & purification; D015941:Complement Hemolytic Activity Assay; D004591:Electrophoresis, Polyacrylamide Gel; D006183:H-2 Antigens / Q000302:isolation & purification*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008811:Mice, Inbred DBA; D008970:Molecular Weight; D011817:Rabbits
D001798:Blood Proteins; C018386:C4a protein, mouse; D003181:Complement C4; D006183:H-2 Antigens