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Determination of subpicomole levels of NADH and FMN using bacterial luciferase and the liquid scintillation spectrometer

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
39
Issue
2
Identifiers
DOI: 10.1016/0003-2697(71)90434-9
Disciplines
  • Biology
  • Economics

Abstract

Abstract A rapid, economical method is described for determining between 10 femtomoles (10 −14 mole) and 100 nanomoles (10 −7 mole) of NADH or FMN. The technique involves the measurement of light produced by a bacterial luciferase enzyme. This is conveniently done in a modern liquid scintillation spectrometer with the photomultipliers switched out-of-coincidence. Various factors which affect the assay of NADH and FMN in biological samples have been examined in detail. The bacterial luciferase may be coupled to enzymes utilizing or producing NADH or FMN to produce dynamic measurements in situ.

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