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Assay Method Information |
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| | Radiometric Assay |
| Description: | The IC50 values for the most active hits were verified by radiometric assay as follows. A mixture of 15 然 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 蚓. 1.8 無 of 25 然 IPP (1% 1-3H IPP, 15 湣i/mL, American Radiolabeled Chemicals, Inc.) was then added. The reaction was incubated at 37 蚓 for 10 min before quenching with 500 無 saturated NaCl solution. The saline solution was extracted with 500 無 butanol by vortexing, and 300 無 of the organic layer was transferred into scintillation vial for radiation readout. |
| Affinity data for this assay | |
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