| Assay Method Information | |
| | Activity Inhibition Assay |
| Description: | Specifically, in order to determine IC50 of the compounds for neuraminidase, 0.01 U/ml of neuraminidase (EC. 3.2.1.8, C. perfringens, SIGMA, N2876) as an enzyme, 4-methylumbelliferyl-a-D-N-acetylneuraminic acid (SIGMA, M8639) mixed in 0.1 mM acetate buffer [50 mM sodium acetate (pH 5.0)] as a substrate, and inhibitors dissolved in methanol in each concentration were prepared. Fluorescence analysis was observed by SpectraMax M3 through excitation at 365 nm and emission at 450 nm (gain 40 nm). The activity analysis was performed on a 96 well plate (SPL Life Sciences, Korea) by providing total 200 μl of a solution comprising an enzyme (10 μl), a substrate (20 μl), an inhibitor (10 μl), and an acetate buffer (160 μl). By plotting initial rates of inhibition reactions occurring at different concentrations of inhibitors, the inhibition capability was evaluated. |
| Affinity data for this assay | |
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