| Assay Method Information | |
| | Inhibitory Activity of Compounds on HDAC1, 2 and 3 |
| Description: | a) Formulation of 100% solution: 50 μL of HDAC buffer was mixed with 10 μL of enzyme solution, 40 μL of substrate was added after 5 minutes and the above materials reacted at 37° C. for 30 minutes, then 100 μL of trypsin termination solution was added to terminate the above reaction, and the reaction was carried out at 37° C. for 20 minutes, the fluorescence intensity was measured at 390 nm/460 nm to obtain 100% absorbence. AMC was used as the standard to create a standard curve and calculate enzyme activity.b) Formulation of blank solution: 60 μL of HDAC buffer was added with 40 μL of substrate, and the above materials reacted at 37° C. for 30 minutes, then 100 μL of trypsin termination solution was added and the reaction was carried out at 37° C. for 20 minutes, the fluorescence intensity was measured at 390 nm/460 nm to obtain blank absorbence.6. The determination steps for drug inhibition of HDAC enzyme activity: 50 μL of HDAC buffer containing a drug was mixed with 10 μL of enzyme solution and pre-incubated for 5 minutes, 40 μL of substrate was added and then the above materials reacted at 37° C. for 30 minutes, then 100 μL of trypsin termination solution was added to terminate the above reaction, and the reaction was carried out at 37° C. for 20 minutes, and the fluorescence intensity was measured at 390 nm/460 nm. |
| Affinity data for this assay | |
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