| Assay Method Information | |
| | Enzyme Activity (EC50) Analysis Assay |
| Description: | In order to confirm the PPARδ activation effect of the example compounds, the following experiment was performed. First, the example compounds were diluted in DMSO to 100× based on the final concentration, then 1/50 was diluted in assay buffer, and 10 μL of each was added to a 384 well plate. Afterwards, GST-conjugated PPARδ ligand-binding domain (LBD) was added to a final concentration of 5 nM, and fluorescien-conjugated C33 coactivator peptide and Tb-α-GST antibody were added to a final concentration of 100 nM and 10 nM, respectively. The final volume of each test system was set to 20 μL, and the test for each concentration was repeated twice, and the binding activity after reaction was measured by TR-FRET method. In other words, after excitation at 340 nm and measurement of emission values at 495 nm and 520 nm, the results were analyzed using the measured value at 490 nm/measured at 520 nm, and the analysis program Prism 6 was used to calculate the EC50 value. |
| Affinity data for this assay | |
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