D2197

Diaphorase from Clostridium kluyveri

recombinant, expressed in E. coli, Animal-component free

• Synonym(s): Diaphorase, Lipoamide Dehydrogenase, Lipoyl Dehydrogenase
• EC Number: 1.8.1.4 (BRENDA, IUBMB)
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• recombinant: expressed in E. coli
• Quality Level: 200
• form: solid
• specific activity: >=30 units/mg protein (biuret)
• storage temp.: −20°C

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.

Unit Definition

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.

Physical form

Supplied as a lyophilized powder containing trehalose.

Safety Information

• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable