| Assay Method Information | |
| | Inbinition of the Menin-MLL Binding |
| Description: | Test for Evaluating the Inbinition of the Menin-MLL BindingMenin1-615 wherein 6× His tag and HA tag are inserted in the N-terminus, and myc tag is inserted in the C-terminus (hereinafter, referred to as His-Menin1-615), was diluted with an assay buffer (25 mmol/L HEPES, 150 mmol/L NaCl, 1 mmol/L dithiothreitol, 0.5% (w/v) Tween 80, 0.3% (w/v) BSA, 0.3% (w/v) skim milk) to adjust the final concentration Lo 30 nmol/L. The test compounds were also diluted with the assay buffer to adjust each concentration of the test compounds to 0.005 to 5 μmol/L. The prepared His-Menin1-615 and test compounds were added to a light-shielding 384-well low-volume plate (Corning, #4514) in 2 μL/well and 6 μL/well, respectively, and the plate was covered with a lid for light-shielding (Corning, r-3935), and incubated at room temperature for 3 hours. After the incubation, MLL1-172 wherein FLAG tag is inserted in the C-terminus (MLL1-172-FLAG), was separately diluted with the assay buffer to adjust the final concentration to 50 nmol/L. The prepared MLL1-172-FLAG was added to the above plate in 2 μL/well, and the plate was covered with a lid for light-shielding and incubated at room temperature for an hour. |
| Affinity data for this assay | |
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