Bioluminescent assay of NADPH-dependent isocitrate dehydrogenase and its substrates and cofactors.
A bioluminescent assay for NADPH-dependent isocitrate dehydrogenase and for its substrates and cofactors was developed. The method is based on continuous NADPH monitoring in the reaction. The linear range of the assay for the enzyme activity is from 0.05 U/liter to 30 U/liter. It is about 300 times more sensitive than the corresponding spectrophotometric assay at 340 nm. Good correlation exists between both assays. Isocitrate, NADP, manganese, and magnesium can be measured at picomole levels. The applicability of the assays to serum analysis is discussed.
D003067:Coenzymes / Q000032:analysis*; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007521:Isocitrate Dehydrogenase / Q000032:analysis*; D007523:Isocitrates / Q000032:analysis; D008163:Luminescent Measurements; D008274:Magnesium / Q000032:analysis; D008345:Manganese / Q000032:analysis; D009249:NADP / Q000032:analysis / Q000502:physiology; D013056:Spectrophotometry, Ultraviolet
D003067:Coenzymes; D007523:Isocitrates; D008345:Manganese; D009249:NADP; C034219:isocitric acid; D007521:Isocitrate Dehydrogenase; D008274:Magnesium