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从牛脑制备NSE纯品.纯化步骤依次为硫酸铵分级沉淀,DEAE—Sephacel离子交换层析,Sephacryl—s—300凝胶过滤层析和DEAE—Sephadex A—50离子交换层析,所得NSE 纯品,PAGE呈单一区带,SDS—PAGE 亚基分子量46KD,IEF—PAGE pI 4.7～4.9,同工酶电泳仅显示γγ酶活性区带,酶比活性30.1 U/mg 蛋白,得率38.6%.
Pure NSE was prepared from bovine brain, followed by ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography, Sephacryl-s-300 gel filtration chromatography and DEAE-Sephadex A-50 ion exchange chromatography. The PAGE and PAGE showed a single band. The molecular weight of SDS-PAGE subunit was 46KD and IEF-PAGE pI was 4.7-4.9. Isoenzyme electrophoresis showed only γγ enzyme activity band with enzyme specific activity of 30.1 U / mg protein, with a yield of 38.6%.