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| | NMR Titration Experiment |
| Description: | NMR measurements consisted of monitoring changes in chemical shifts and line widths of the backbone amide resonances of uniformly 15N-enriched MDM2 samples in a series of HSQC spectra as a function of a ligand concentration. The complexes of human MDM2 and the chalcones were prepared by mixing the protein and the ligand in the NMR tube. Typically, NMR spectra were recorded 15 min after mixing at room temperature. An initial screening of all compounds used in this study was performed with a 10-fold molar excess of chalcone to human MDM2. All subsequent titrations were carried out until no further shifts were observed in the spectra. All KD values obtained by NMR spectroscopy are based on at least six data points. |
| Affinity data for this assay | |
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