| Assay Method Information | |
| | SAMR1 (50-724) Enzymatic Assay |
| Description: | The enzymatic assay was performed in a 384-well plate using Dulbecco's PBS buffer as the reaction buffer. Purified SARM1 (50-724) with a final concentration of 2 nM was pre-incubated with the test compound at 1% DMSO final assay concentration for 15 min at room temperature. The reaction was initiated by adding the mixture of 200 μM NMN as activator and 100 μM NAD+ as substrate. After 1 h of incubation at room temperature, the reaction was terminated with 10 times volume of 70% acetonitrile, then centrifuged at 3800 rpm for 10 min. The samples were analyzed using LC-MS/MS after diluted to the proper concentration by 10 mM ammonium acetate (pH 9.75). |
| Affinity data for this assay | |
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