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Hydrogen peroxide (HOOH) detection assay for LSD1. When a dimethylated lysine substrate (bottom left) is demethylated by LSD1, the FAD cofactor is reduced to FADH2 (center). The cofactor is reoxidized by molecular oxygen to form HOOH (top center). Horseradish peroxidase (HRP) uses this to couple 4-aminoantipyrine and dichlorohydroxybenzenesulfonate (top right) to form the chromophore (center right) that is detected at 515 nm (bottom right).
