Bioluminescent assay of lactate dehydrogenase and its isoenzyme-1 activity.
A bioluminescent assay based on the bacterial luciferase reaction has been developed for the determination of total lactate dehydrogenase and heart-specific lactate dehydrogenase isoenzyme-1 activity in serum. The lactate dehydrogenase-catalyzed reaction was measured in both directions, but NADH formation (lactate----pyruvate) is recommended because it allows the use of optimal reaction conditions. Internal calibration with a known amount of NADH accounts for possible interference from samples when both NADH formation and consumption are followed. The bioluminescent method is sensitive, has good precision, and is readily automated. Serum lactate dehydrogenase isoenzyme-1 was immunochemically isolated and the activity was assayed by bioluminescence. A good correlation between the bioluminescent assays and the conventional spectrophotometric procedure used as reference was obtained.
D006801:Humans; D007527:Isoenzymes; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000097:blood*; D008156:Luciferases; D008163:Luminescent Measurements; D008722:Methods; D009206:Myocardium / Q000201:enzymology; D009243:NAD / Q000032:analysis; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet
D007527:Isoenzymes; D009243:NAD; D007770:L-Lactate Dehydrogenase; D008156:Luciferases