title: Biochemical and genetic analysis of NAD(P)H:quinone oxidoreductase 1 (NQO1).
abstract: NAD(P)H:quinone oxidoreductase 1 (NQO1, DT-diaphorase, E.C. 1.6.99.2) is an FAD containing obligate two-electron reductase that catalyzes the reduction of a broad range of substrates. This unit will describe methods for the detection of NQO1 protein in formalin-fixed, paraffin-embedded tissues by immunohistochemistry; detection of NQO1 protein in fresh tissues or cell lines by immunoblot analysis; measurement of NQO1 catalytic activity in fresh and frozen tissues and cell lines using spectrophotometric assays based upon the reduction of 2,6-dichlorophenol-indophenol (DCPIP) or coupled menadione-cytochrome reduction; and determination of the NQO1*2 polymorphism by PCR-RFLP.
authors.affiliation: University of Colorado at Denver and Health Sciences Center, Denver, Colorado, USA.
authors.forename: David
authors.initials: D
authors.lastname: Siegel
authors.forename: Jadwiga K
authors.initials: JK
authors.lastname: Kepa
authors.forename: David
authors.initials: D
authors.lastname: Ross
mesh_terms: D001483:Base Sequence; D002384:Catalysis; D002460:Cell Line; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013053:Spectrophotometry
chemical_list: D004364:Pharmaceutical Preparations; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human