title: Extraction and measurement of NAD(P)(+) and NAD(P)H.
abstract: Nicotinamide adenine dinucleotides are critical redox-active substrates for countless catabolic and anabolic reactions. Ratios of NAD(+) to NADH and NADP(+) to NADPH are therefore considered key indicators of the overall intracellular redox potential and metabolic state. These ratios can be measured in bulk conditions using a highly sensitive enzyme cycling-based colorimetric assay (detection limit at or below 0.05 μM or 1 pmol) following a simple extraction procedure involving solutions of acid and base. Special considerations are necessary to avoid measurement artifacts caused by the presence of endogenous redox-active metabolites, such as phenazines made by diverse Pseudomonas species (see Chapter 25).
authors.affiliation: Biology Department, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: Suzanne E
authors.initials: SE
authors.lastname: Kern
authors.forename: Alexa
authors.initials: A
authors.lastname: Price-Whelan
authors.forename: Dianne K
authors.initials: DK
authors.lastname: Newman
mesh_terms: D001671:Biochemistry / Q000379:methods*; D009249:NADP / Q000032:analysis* / Q000302:isolation & purification*; D011549:Pseudomonas / Q000378:metabolism; D012015:Reference Standards; D013223:Statistics as Topic
chemical_list: D009249:NADP