| Assay Method Information | |
| | Radiometric Assay |
| Description: | The IC50 values for the most active hits were verified by radiometric assay as follows. A mixture of 15 µM cis-FPP, 100 nM DPPS, and inhibitors in the assay buffer (25 mM Tris-HCl, 1 mM MgCl2, 0.01% Triton-X-100) was incubated for 10 min at 25 °C. 1.8 µL of 25 µM IPP (1% 1-3H IPP, 15 µCi/mL, American Radiolabeled Chemicals, Inc.) was then added. The reaction was incubated at 37 °C for 10 min before quenching with 500 µL saturated NaCl solution. The saline solution was extracted with 500 µL butanol by vortexing, and 300 µL of the organic layer was transferred into scintillation vial for radiation readout. |
| Affinity data for this assay | |
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