| Assay Method Information | |
| | Menin-MLL Interaction Inhibitory Activity Assay |
| Description: | The inhibition of Menin/MLL-4-43 peptide interaction by small molecule inhibitors was quantitatively detected by fluorescence polarization competition assay. The experiment was performed in a 384-well plate (Corning, Cat #3575) using a reaction buffer consisting of 50 mM Tris, pH 7.5, 50 mM NaCl, and 1 mM DTT. A 40 μL reaction system included 10 μL of 8 nM Menin recombinant protein and 10 μL of the test compound at different concentrations. The compound was pre-incubated with Menin protein for 15 min, and then 20 μL of 10 nM FITC-MLL4-43-peptide was added. After incubation on a shaker at 25° C. for 60 min, the fluorescence polarization signal (FP 485 520 520) was read using BMG PHERAStar detection. The experimental data was analyzed and processed by GraphPad Prism 6 software to obtain the IC50 value. |
| Affinity data for this assay | |
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