| Assay Method Information | |
| | HDAC Inhibition Assay |
| Description: | HDAC activity was measured using a continuous trypsin-coupled assay. For inhibitor characterization, measurements were done using 96-well assay plates. Enzyme was mixed with inhibitor at various concentrations and allowed to incubate for 15 minutes. Trypsin was added to initiate the reaction. Reactions were monitored in a fluorescence plate reader. After a 30-minute lag time, the fluorescence was measured over a 30-minute time frame using an excitation wavelength of 355 nm and a detection wavelength of 460 nm. The increase in fluorescence with time was used as the measure of the reaction rate. Inhibition constants Ki(app) were obtained using the program BatchKi. |
| Affinity data for this assay | |
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