| Assay Method Information | |
| | Inhibition of Fabl Protein |
| Description: | Inhibition of Fabl enzyme from Acinetobacter baumannii and Escherichia coli was tested by measuring the rate of NADH consumption (Aabsorbance at 340 nm/min) at 30° C. in 96-well plate format using an automated plate reader in the presence or absence of the test compounds. The assay mixture contained 100 mM Tris-HCl, pH 7.25 (A. baumannii) or 7.5 (E. coli), 100 mM ammonium acetate, 0.02% (A. baumannii) or 0.05% (E. coli) Pluronic F-68, 25 μM crotonyl ACP, 50 μM NADH, 25 μM (A. baumannii) or 50 μM (E. coli) recombinant Fabl protein, and 7.5% DMSO. Test compounds were added at concentrations ranging from 0.17 to 10,000 nM in a final well volume of 100 μl. This dose-response inhibitory assay was performed using a 10-point, serial dilution series for each test compound. IC50 values for each test compound were assigned from logistical sigmoid curve-fitting of the inhibition dose response curves. |
| Affinity data for this assay | |
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