| Assay Method Information | |
| | NAD+ Cydase Activity Assay with Human or Mouse Recombinant CD38 Protein |
| Description: | Required reaction buffer, required CD38 concentration in reaction buffer and required e-NAD concentration in reaction buffer were prepared. Thereafter, 8 μL of CD38 in reaction buffer were added to High Control and compound concentration wells in 384 well plate (Corning #4514, 384 well, nonbinding surface, low volume, round bottom) and 8 μL of reaction buffer to Low Control. Compound dilutions were transferred via pintool technology that transfers 42.5 nL compound dilutions (12-point dose-response with factor three dilutions in 100% DMSO) well-to-well. Plate was subsequently shaken for 15 s at 1450 rpm and sealed for incubation for 30 min at room temperature. Thereafter, 2 μL of e-NAD in reaction buffer were added with Integra Voyager. After shaking for 15 s at 1450 rpm, plate was sealed, and assay signal was measured continuously with PHERAstar® fluorescence intensity module (extinction 300 nm/emission 410 nm). |
| Affinity data for this assay | |
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