| Assay Method Information | |
| | In Vitro ANT(2")-Ia Pyrophosphatase Assay |
| Description: | In vitro ANT(2")-Ia pyrophosphatase assays were performed as described previously (Cox et al., 2015; Hirsch et al., 2014), with minor modifications. To improve compound solubility, Tween 20 was added to a final concentration of 0.01% (v/v) and compounds were added at a final concentration of 2% (v/v) DMSO. The enzyme (1 mg per 100 mL reaction volume) was incubated with compound for 10 min at 25 °C, prior to initiation of the reaction with nucleotide. All inhibitory assays were performed with kanamycin B kept at Km (Cox et al., 2015; Hirsch et al., 2014). |
| Affinity data for this assay | |
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