Description
General description
Research area: Cancer
Application
Diaphorase from Clostridium kluyveri has been used:in lactate dehydrogenase assay in gill cells.
impregnated on the strip for formate dehydrogenase assay.
in various assays to oxidate β-NADH or β-NADPH in the presence of an electron acceptor.
for semiquinone generation during benzoquinone reduction.
for evaluation of glucose uptake in cultured muscle cells.
in lactate dehydrogenase cytotoxicity assay.
Biochem/physiol Actions
Diaphorase catalyzes the oxidation of either β-NADH or β-NADPH in the presence of an electron acceptor such as methylene blue or 2,6-dichlorophenolindophenol. Diaphorases that are specific for either β-NADH or β-NADPH are known. The pig heart enzyme of Straub seems to have native diaphorase (β-NADH specific) as well as lipoic and lipoamide dehydrogenase activities.
Physical form
Supplied as a lyophilized powder containing trehalose.
Other Notes
One unit of diaphorase will oxidize 1.0 micromole of beta-NADH per minute at pH 7.5 at 25 degrees C, with the corresponding reduction of the appropriate electron acceptor.