| Assay Method Information | |
| | Inhibition Kinetics Assay |
| Description: | Slow-onset inhibition kinetics were monitored at 340 nm on a Cary 100 spectrophotometer (Varian) at 25 °C in 30 mM PIPES buffer (pH 8.0) containing 150 mM NaCl and 1.0 mM EDTA. The reactions were initiated by the addition of enzyme (8 nM) to a mixture containing glycerol [8% (v/v)], bovine serum albumin (0.1 mg/mL),DMSO [2% (v/v)], crot-CoA (750 μM), NADH (250 μM), NAD+ (200 μM), and inhibitor (0−8000 nM). All reactions were monitored until the steady state was reached, indicated by the linearity of the progress curve. |
| Affinity data for this assay | |
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