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Flavodoxin-like proteins (Fld-LPs) are an important constituent of the oxidative stress defense system in several organisms and highly conserved from bacteria to humans. These proteins possess NAD(P)Hquinone oxidoreductase activity and convert quinones to hydroquinones through two-electron reduction, using NAD(P)H and quinone as electron donor and acceptor, respectively. Purified yeast and bacterial Fld-LPs exhibit NAD(P)Hquinone oxidoreductase activity in vitro. Here, we describe a protocol to measure oxidoreductase activity of Fld-LPs t