Optimal conditions for assaying human lactate dehydrogenase by the lactate-to-pyruvate reaction: Arrhenium relationships for lactate dehydrogenase isoenzymes 1 and 5.
Optimal reaction conditions to sassay human lactate dehydrogenase (lactate-to-pyruvate) were established for isoenzymes 1 and 5 at 25, 30, and 37 degrees C in diethanolamine and 2-amino-2-methyl-1,3-propanediol. Different substrate concentrations are required at each temperature. The conditions permit measurement of lactate dehydrogenase 1 and 5 with the lowest substrate concentrations that allow for the highest equal sustainable efficiency in measuring both isoenzymes. About 95% of each isoenzyme activity is measured if the assay is performed within the first minute after the reaction is initiated even for activities as high as triple the upper limit of normal. The Arrhenius relationship is different for each isoenzyme, but results obtained for each at one temperature can be compared with results at another temperature by use of simple conversion equations. Assays at 25 and 30 degrees C are more economical and less variable than assays at 37 degrees C.
D002021:Buffers; D006801:Humans; D066298:In Vitro Techniques; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000032:analysis* / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D008722:Methods; D009243:NAD; D011773:Pyruvates / Q000096:biosynthesis; D013696:Temperature; D013816:Thermodynamics
D002021:Buffers; D007527:Isoenzymes; D007773:Lactates; D011773:Pyruvates; D009243:NAD; D007770:L-Lactate Dehydrogenase