Purification of creatine kinase-BB isoenzymes from human and canine tissues

We describe a procedure for the purification of creatine kinase BB isoenzyme from canine and human brain tissue. The purification involves sequentially 50-70% (NH4)2SO4 fractionation, DEAE-Sephacel, Blue-Sepharose CL-6B, and chromatofocusing chromatography. This method produces a homogeneous protein purification as verified by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing chromatography.