| Assay Method Information | |
| | Fluorescence-Based PFTase Inhibition Assay |
| Description: | Assays were realized on 96-well plates, prepared with Biomek NKMC and Biomek 3000 from Beckman Coulter and read on a Wallac Victor fluorimeter from PerkinElmer. Per well, 20 μL of farnesyl pyrophosphate (10 μM) was added to 180 μL of a solution containing 2 μL of varied concentrations of tested compounds (dissolved in methanol) and 178 μL of a solution composed by 0.1 mL of partially purified recombinant yeast FTase (2.2mg/mL) and 7.0 mL of Dansyl-GCVLS peptide (in the following buffer: 5.8 mM DTT, 12 mM MgCl2, 12 μM ZnCl2, and 0.09% (w/v) CHAPS, 53 mM Tris/HCl, pH 7.5). Then the fluorescence development was recorded for 15 min (0.7 s per well, 20 repeats) at 30°C with an excitation filter at 340 nm and an emission filter at 486 nm. |
| Affinity data for this assay | |
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