| Assay Method Information | |
| | FAS Activity Assay |
| Description: | FAS activity was determined by monitoring the decrease in absorbence at 340 nm resulting from the oxidation of NADPH using an Amersham Pharmacia Ultrospec 4300 pro UV-Vis spectrophotometer at 37°C. The assay mixture contained 100 mM potassium phosphate buffer, pH 7.0, 1 mM EDTA (ethylenediaminetetraacetic acid), 2.5 μM acetyl-CoA, 10 μM malonyl-CoA, 32 μM NADPH, and 10 μg chicken liver FAS in a total volume of 2.0 mL [Tian et al., J. Biol. Chem., 260:11375-87]. Fast inhibition was determined by adding thioether to the reaction system before FAS initiated the reaction. The initial velocity was measured to calculate the remaining activity (RA) of FAS, and each assay was repeated three times. |
| Affinity data for this assay | |
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