| Assay Method Information | |
| | NSDHL Inhibitory Activity Evaluation |
| Description: | An activity assay was performed using a 96-well plate, and an experiment was performed after adding an inhibitor to each well at the final concentrations of 200 μM≤1 μM. Particularly, 50 μl of 16 μM NSDHL, 50 mM HEPES, 20% glycerol were added to each well, and then 50 ul of 50 μM NADH was added to each well, followed by reaction at room temperature for 10 minutes. The fluorescence level of each well was measured using SpectraMax M5 (Molecular Devices) equipment (Ex: 340 nm, Em: 460 nm). The values were corrected by separately measuring the absorbance of the compound and the buffer solution, and the degree of inhibition of each compound against NSDHL was measured based on the NADH fluorescence at the same concentration without binding. IC50 (inhibitory concentration at 50%) was calculated using GraphPad Prism software. |
| Affinity data for this assay | |
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