| Assay Method Information | |
| | In Vitro hDDAH-1 Assay |
| Description: | A colorimetric assay was carried out in 150 μL 50 mM potassium phosphate buffer pH 7.4 containing 4 μg of recombinant hDDAH-1 and varying concentrations of NMMA and inhibitor. NMMA was applied at 50, 100, 300, 700 and 1250 μM. Each dilution was incubated each inhibitor in five different concentrations and one without inhibitor as a control. Samples were incubated in a 96-well microplate at 37°C for 30 min and the reaction was stopped by addition of 200 μL Colder reagent [Knipp et al., Anal. Biochem., 286:257-264]. Microplates were sealed with sealing tape, incubated at 95°C for 20 min and then read at 540.5 nm. |
| Affinity data for this assay | |
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