| Assay Method Information | |
| | TryR Enzyme Inhibition Assay |
| Description: | Inhibition of TryR was carried out in 96-well plates using a Biotek Precision 2000 automated liquid handler. Reaction was initiated by addition of NADPH to the assay mixtures contained TryR , trypanothione disulfide substrate, and inhibitor (100 uM to 10 nM in 3-fold serial dilutions). The linear rate of thionitrobenzoate ion formation was monitored over 5 min in a Spectramax 340PC plate reader (Molecular Devices) at 412 nm. Raw data was processed using Microsoft Excel. Grafit 5.0 (Erithacus software) was used to fit the data to a three-parameter equation to determine IC50 values. IC50 determinations were carried out in triplicate for each compound and the mean weighted to standard error calculated. |
| Affinity data for this assay | |
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