| Assay Method Information | |
| | Enzyme Inhibitory Efficacy Assay |
| Description: | Specific experimental methods are as follows. First, the compounds of Examples 1 to 89 of 1 nM to 100 μM (4 conc.) were put into a 384 well plate (25 μL/well), and 2 μM EEMT2/G9a enzyme (4 conc.) was put into the 384 well plate (25 μL/well). The plate was then centrifuged at 1,500 rpm for 5 seconds after sealing the plate with plate sealing foil. After incubating the EHMT2/G9a enzyme, SAM, and the compounds of Examples above at room temperature for 10 minutes, 2 μM histone H3 peptide (4 conc.) was added to the 384-well plate (25 μL/well). After sealing the plate with plate sealing foil and centrifuging at 1,500 rpm for 5 seconds, an enzymatic reaction was performed at room temperature for 1 hour. Upon completion of the enzyme reaction, 5 μL each of 200 nM Ulight -Streptavidin (4 conc.) and 8 nM Eu-antibody (4 conc.) was added to each well of the plate. The plate was sealed with plate sealing foil, centrifuged at 1,500 rpm for 5 seconds and reacted at room temperature for 1 hour. The signal values were measured with Envision (Ex 320 nm, Em 665 nm), and all background values obtained by reacting without enzyme were deleted from the measured raw data values. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |