Purification andkinetic mechanism ofthemajor glutathione S-transferase frombovine brain

toproceed for 20h at 37°C in a shaking incubator. Thecoupled gel was washed and remaining groups wereblockedbythe addition of1.0 M-ethanolamine. After4hthegelwaswashedwith(1 litre each)0.5M-KCIin0.10M-sodium acetate buffer, pH4.0, 0.10 M-sodium boratebuffer, pH8.0, and finally with 0.10 M-potassiumphosphate buffer, pH8.0. Thegel waspoured in a glasscolumn (0.5 cmx 10cm). In general, up to 30mg ofDEAE-cellulose-purified protein could beapplied to thecolumn with little loss in separating efficiency. Proteinwas applied in 0.02M-potassium phosphate buffer,pH7.0, containing