| Assay Method Information | |
| | FAS Inhibitory Assay |
| Description: | To each microtube (final volume: 100 μL), FAS was added (20-30 μg protein) in a buffer containing 100 mM potassium phosphate (pH 7.0), 2.5 mM dithiothreitol and 2.0 mM EDTA with or without test compounds. The mixtures were preincubated at 37°C for 60 min and the reaction was started by the addition of a substrate mixture containing 0.25 mM NADPH, 0.4 nmol of malonyl CoA and 0.02 μCi of [3H] acetyl CoA. Following incubation at 37°C for 10 min, the reaction was terminated with 60% HClO4. Fatty acids were extracted with 1 mL of hexane and incorporation of radioactivity into the fatty acids was assessed by scintillation counting [Na et al., Bioorg. Med. Chem. Lett., 16:4738-4742]. |
| Affinity data for this assay | |
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