| Assay Method Information | |
| | Measurement of NamPT Inhibitory Ability IC50 |
| Description: | To test the effect of the compound prepared in each Example on the substrate decomposition rate (Vmax/min) mediated by NamPT enzyme, IC50 was calculated. In this experiment, the NamPT colorimetric assay kit (ab221819) produced by Abcam was used, and Graph prism software was used for IC50 calculation. PRPP, ATP, NMNAT1, NamPT recombinant proteins, and various concentrations of the compound of each Example (half-log step dilution from 100 μM) were added to nicotinamide to synthesize nicotinamide adenine dinucleotide (NAD) from the nicotinamide, and a total of 60 μL of the reaction solution was reacted at room temperature for 1 hour. WST-1 formazan converted by NAD generated by adding WST-1 was measured. The result of the reaction was obtained by the following formula by measuring absorbance using EnVision2105 (PerkinElmer) at 450 nm for 60 minutes at 2-minute intervals. |
| Affinity data for this assay | |
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