Wallac Oy

Jukka Lavi

Timo Lovgren

A method for determining NADH and/or NADPH-concentrations, for instance in NADH- or NADPH transforming systems, the method comprising the step of bringing the sample subject to determination in contact with the bioluminescent reagent based on bacteria luciferase, NAD(P)H-FMN oxidoreductase, FMN and aliphatic aldehyde, whereby a reaction takes place, where NAD(P)H is oxidized and FMN is reduced, the reaction being catalyzed by the oxidoreductase, whereafter the FMNH.sub.2 formed and the aldehyde are bound to the luciferase and light is emitted, the intensity of this light being measured and constituting a measure of the NADH- or NADPH-concentration, whereby in the determination a bacteria luciferase purified so as not to obtain any oxidoreductase is used in combination with a predetermined amount of oxidoreductase, this amount being such that a stable light intensity is obtained in the presence of a constant amount of NADH or NADPH whereby an internal calibration can be carried out in NADH or NADPH transforming systems by adding a predetermined amount of NADH or NADPH followed by a subsequent measuring of the increase of the light intensity.

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Karjatie, Finland

Jukka Lavi