| Assay Method Information | |
| | Fluorescence Polarization (FP) Assay |
| Description: | Compounds were tested in a biochemical binding assay using Menin protein at 2 nM (N6his-tev-Menin_4i80, PB-20-1459) and the substrate cRho110-Ahx-MBM1/MBM2 peptide (UbiQ, UbiQ-Q20201030; cRh110-Ahx-SCRWRFPARPGTTGGGGGGGRRGLGGAPR-QRVPALLLPPG-NH2) at 1 nM.384 Low volume black plates (Corning #4514) were used. 5 μL/well compound or 3% DMSO was added at 10-fold serial dilutions in DMSO from 10 μM. 5 μL/well cRho110-Ahx-MBM1/MBM2 peptide or binding buffer (50 mM Tris pH 7.5, 50 mM NaCl, 1 mM TCEP, 0.01% BGG, 0.01% Brij-35) was added to all wells. 5 μL/well Menin protein or binding buffer (50 mM Tris pH 7.5, 50 mM NaCl, 1 mM TCEP, 0.01% BGG, 0.01% Brij-35) was added to all wells. Plates were read on an EnVision plate reader (Perkin Elmer) with excitation at 480 nm and emission at 535 nm P+S, and measured every 5 minutes for 180 minutes. |
| Affinity data for this assay | |
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