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Assay Method Information |
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| | Diaphorase/Resazurin Coupled Assay |
| Description: | For WT IDH1 and IDH2, reactions were conducted at room temperature in 384-well Greiner black microtiter plates in a total volume of 10 μL of assay buffer. Final compound concentrations were typically varied from 5 to 100,000 nM; the isocitrate concentration was fixed at 10 μM, the NADP+ concentration was fixed at 5 μM, and WT IDH1 and IDH2 were fixed at 0.1 nM. Reactions were conducted in duplicate and run kinetically. After each addition, plates were centrifuged for 60 s to ensure complete mixing of reagents. |
| Affinity data for this assay | |
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