The purification of human creatine kinase BB with high specific activity.
This paper describes the purification of human creatine kinase BB with high specific activity (1,122 U/mg). The procedure used resulted in a protein yield of 5.4 mg (21% recovery) from 150 g of brain tissue. Two-dimensional electrophoresis and PAGE studies indicated that purified CK-BB might exist as native isoenzyme along with structural aggregates since the multi-banded appearance was reduced to a single band with sodium dodecyl sulfate treatment but not with 2-mercaptoethanol. Investigators are cautioned not to store brain tissue for prolonged periods of time before isolation of the isoenzyme as this may lead to protein redistribution with additional bands becoming evident on PAGE.
D001921:Brain / Q000201:enzymology*; D002850:Chromatography, Gel; D003402:Creatine Kinase / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004589:Electrophoresis, Disc; D006801:Humans; D007525:Isoelectric Focusing; D007527:Isoenzymes; D008623:Mercaptoethanol; D008970:Molecular Weight; D013048:Specimen Handling; D013194:Staining and Labeling
D007527:Isoenzymes; D008623:Mercaptoethanol; D003402:Creatine Kinase