Related Experiment Video
Updated: Jun 21, 2026

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
Published on: February 24, 2018
Measurement of cytochrome P450 and NADPH-cytochrome P450 reductase
F Peter Guengerich1, Martha V Martin, Christal D Sohl
1Department of Biochemistry and Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee, USA. f.guengerich@vanderbilt.edu
Abstract:
Cytochrome P450 (P450) enzymes are important in the metabolism of steroids, vitamins, carcinogens, drugs and other compounds. Two of the commonly used assays in this field are the measurements of total P450 and NADPH-P450 reductase in biological preparations. A detailed protocol is presented for the measurement of P450 by its spectral properties, along with a protocol for measuring NADPH-P450 reductase by its NADPH-cytochrome c reduction activity. Each assay can be completed in 5-10 min. Detailed explanations for the rationale of particular sequences in the protocols are provided, along with potential confounding problems.
Related Concept Videos
Role of Reduced Coenzymes NADH and FADH₂
Redox Reactions
Phase I Reactions: Reductive Reactions
Pharmacogenetics of Phase I Enzymes: Cytochrome P450 Isozymes

