| Assay Method Information | |
| | Evaluation of Enzyme Activity |
| Description: | Table 1: The purpose of the assay was to test the in vitro inhibitory activity of the compounds against LSD1. The enzyme used in the assay was human LSD1, and the standard substrate was histone H3K4me peptide (20 μM). The activity of the compounds was determined by the enzyme-coupling fluorescent method by the combined detection of H2O2 generated after the reaction of LSD1 by horseradish peroxidase (HRP) and the fluorescent reagent Amplex Red. Starting from 10 μM, the compounds were 3-fold diluted to detect the IC50 values of the compounds at 10 concentrations. The enzyme and substrate were incubated for 30 minutes before the compound was added to the substrate to start the reaction. Fluorescence detector: EnVision, excitation wavelength: Ex/Em=530/590 nM. |
| Affinity data for this assay | |
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