| Assay Method Information | |
| | Test on EZH2 Inhibitory Activity |
| Description: | Table 1: Test ConditionsEZH2 enzyme complex: 25 ng/uLHistone H3 peptide substrate: 4 uMSAM: 2 uMReaction time: 300 minProcedures1. Each compound disclosed herein was diluted with 1×HMT buffer and transferred to a 384-well assay plate at 3 uL per well;2. The EZH2 enzyme solution was prepared with 1×HMT buffer and transferred to the 384-well assay plate at 2 uL per well;3. The above assay plate was incubated for 30 min;4. A mixture solution of SAM and histone H3 peptide substrate at 2-fold concentration was prepared with 4×HMT buffer;5. 5 uL of the mixture solution of SAM and substrate was added to each well of the assay plate to react at room temperature, and the assay plate was sealed with plastic;6. After incubation for 5 h, 10 uL of TR-FRET detection buffer containing Tb-labeled antibody and dye-labeled receptor was added to each well of the assay plate;7. After incubation for 1 h at room temperature, the assay plate was detected;8. Data were processed using Prizm software. |
| Affinity data for this assay | |
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