| Assay Method Information | |
| | Formaldehyde Dehydrogenase-Coupled Demethylase (FDH) Assay |
| Description: | For inhibition assays, powders of inhibitor compounds were dissolved in 100% (v/v) DMSO to give a 20 or 50 mM concentration and stored at -20 °C until needed. Various compounds with the concentrations ranging from 1.25 mM to 5 nM in a half-log serial dilution were pre-incubated with the reaction mixture for 15 min (0.5 mM enzyme [E], 15 mM peptide [S], 50 mM (NH4)2Fe(SO4)2, 0.1 mM αKG, and variable inhibitor [I] in the KDM5 reaction buffer with 10% DMSO). The addition of peptide and APAD+ initiated the reaction for 15 min at room temperature. |
| Affinity data for this assay | |
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