| Assay Method Information | |
| | Inhibitory Effect on ACAT2 Activity |
| Description: | 1. Purpose:The analogues of Pyripyropene A structure were tested for their inhibitory effect on ACAT2 activity at intact cellular level by a method in which a fluorescent-labeled sterol is used for determining ACAT2 activity.2. Principle:An inhibition curve was plotted based on that different concentrations of compounds would cause the fluorescence intensity to change by inhibiting the synthesis of the ester containing NBD22-fluorescent labeled sterol, thereby IC50 was calculated.3. Process:HepG2 cells were cultured in a 96-well plate at a starting density of 1.5×104 cells per well for 24 hours. After the cholesterol mixture was well mixed, the cells were cultured for another 24 hours. Then NBD22-fluorescent labeled sterol at a final concentration of 0.5 μg/ml, and compounds at a final concentration gradient of 0, 0.008, 0.04, 0.2, 1 and 5 μM were added. Three wells were set for each concentration. After incubating for 6 hours, the fluorescence intensity was measured using a fluorescence analyzer (E488, A535). The fluorescence intensity values were plotted against different concentrations of the compounds and IC50 was obtained. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |