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作者用成年大鼠心脏制备线粒体AST(mAST)、用人红血球制备细胞浆AST(cAST)。柱层析根据Sampson等的方法,用低浓度NaCl溶液洗脱时,cAST被DEAE-Sephadex A-50吸附,mAST被洗脱;高浓度时,cAST也被洗脱。第一洗脱液含50 mM/L NaCl、15μM/L磷酸吡哆醛、50mM/L Tris、pH 8.5;将其过柱(微量凝胶柱、床高×直径=10×0.6cm);收集最后1 ml洗脱液单独保存(第一部分),作为酶活性测定的空白管。先后加1 ml标本和5 ml第一洗脱液过柱,收集6 ml溶液,内含mAST(第二部分)。此时,再加1 ml第一洗脱液,收集为第三部分,应不
The authors prepared mitochondrial AST (mAST) from adult rat hearts and prepared human cytoplasm AST (cAST) using human erythrocytes. Column chromatography According to the method of Sampson et al., when eluted with a low concentration of NaCl solution, cAST was adsorbed by DEAE-Sephadex A-50 and mAST was eluted; at high concentrations, cAST was also eluted. The first eluate contained 50 mM/L NaCl, 15 μM/L pyridoxal phosphate, 50 mM/L Tris, pH 8.5; it was passed through the column (microgel column, bed height × diameter = 10 × 0.6 cm); collected The last 1 ml of eluate was stored separately (Part 1) as a blank tube for enzyme activity assay. Add 1 ml of the sample and 5 ml of the first eluent to the column and collect 6 ml of the solution containing mAST (Part 2). At this point, add 1 ml of the first eluent and collect it as the third part, which should not be