Methods in Molecular Biology Volume 962, 2013, pp 95-111

Identification of Small Molecules Affecting p53-MDM2/MDMX Interaction by Fluorescence Polarization

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Abstract

Fluorescence polarization (FP) has become a powerful technique to quantitatively analyze the binding of a small soluble fluorescence-labeled probe to a larger soluble protein and its displacement by other molecules. Here, we describe a detailed protocol to identify small molecules capable of targeting p53-MDM2/MDMX interactions using a fluorescence polarization assay with Rhodamine-labeled p53 peptides.