| Assay Method Information | |
| | NAMPT enzyme assay |
| Description: | The reaction was carried out in Buffer A (50 mM Hepes pH 7.5, 50 mM NaCl, 5 mM MgCl2, and 1 mM THP) in 96-well plates. NAMPT protein was added to 1 Buf 1 µL compound, incubated for 15 minutes at room temperature, and then 10x substrate and co-factors in were added to the test well (final concentrations were 30 nM NAMPT, 1 µM NAM, 100 µM PRPP, and 2.5 mM ATP). The reaction was incubated for 30 minutes at room temperature and then quenched with formic acid and L-cystathionine to make final concentration of 1% formic acid and 10 µM L-cystathionine. |
| Affinity data for this assay | |
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