| Assay Method Information | |
| | IDO Coupled Biochemical Assays |
| Description: | In two separate assays, 0.045 μg/μl of human IDO protein was pre-incubated for 10 min at RT with test compounds NewLink 1 and Incyte 1 in the presence of 50 mM KPO4, pH 7.0, 0.5 mM EDTA, 0.5 mM EGTA, 0.05% Triton X-100, 20 mM ascorbate, 500 U/ml catalase, 10 μM methylene blue at RT in a 384 well plate. 0.05 μg/μl kynurenine formamidase (drosophila) and 45 μM L-tryptophan (L-Trp) were added and the assays were incubated at RT for 17 min. Assays were stopped and the level of kynurenine was determined by incubation with Ehrlich's reagent to a final concentration of 1.33% at RT for 5 min. Fluorescence intensity was read at 475 nm/530 nm. |
| Affinity data for this assay | |
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