| Assay Method Information | |
| | Determination of Inhibition Constants |
| Description: | For determination of the inhibition constants (Ki), enzyme inhibition was studied at three different substrate concentrations and seven different concentrations of the inhibitors. The inhibitor was first mixed with the enzyme and incubated 10 min before the reaction was started by addition of the respective substrate solution. The absorbance was monitored for 150 s at 30 s intervals in a spectrophotometer (Spectra Max Molecular Devices, Sunnyvale, CA). After subtraction of the background (measured at 620 nm) from the signal (measured at 405 nm), the steady-state reaction rates were used for construction of Dixon plots and the Ki value was calculated. |
| Affinity data for this assay | |
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