[Isolation of highly-purified NADP(H)-dependent enzymes from the rat liver using NADP-hydrazidoadipoyl oxypropyl...

NADP-hydrazidoadipoyl oxypropyl sepharose was synthesized from epoxyactivated sepharose through a hydrazid derivative and used for isolation of NADP(H)-dependent enzymes such as glutathione reductase, isocitrate dehydrogenase and malate dehydrogenase. The isolation technique involves fractionation with ammonium sulphate, affinity chromatography on NADP-hydrazidoadipoyl oxypropyl sepharose and chromatography on hydroxylapatite. The proposed technique enabled the authors to obtain malate dehydrogenase isocitrate dehydrogenase and glutathione reductase preparations homogeneous according to SDS-electrophoresis in polyacrylamide gel.