| Assay Method Information | |
| | AICAR Tfase Inhibition Assay |
| Description: | The human ATIC enzyme was used for the inhibition assay. The assay buffer was flushed with nitrogen to minimize oxidization of cofactor 10-f-THF. The reaction was initiated by adding the substrate AICAR to reaction mixtures containing ATIC, test compounds, and different concentrations of cofactor. Using a SpectraMax Plus384 microplate reader, the reaction was monitored at 298 nm by measuring the increase in absorbance corresponding to the formation of THF. The THF was generated in the transfer reaction of the formyl group from cofactor to AICAR to produce 5-formyl-AICAR. IC50 values were measured from dose-response curves. |
| Affinity data for this assay | |
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