Open AccessFluorescence Polarization (FP) Assays for Monitoring Peptide‐Protein or Nucleic Acid‐Protein Binding
Author(s) -
Moerke Nathan J.
Publication year - 2009
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch090102
Subject(s) - fluorescence anisotropy , nucleic acid , chemistry , fluorescence , epitope , peptide , oligonucleotide , protein–protein interaction , ligand binding assay , small molecule , biochemistry , biophysics , biology , antigen , dna , receptor , genetics , physics , quantum mechanics , membrane
Abstract The technique of fluorescence polarization (FP) is based on the observation that when a fluorescently labeled molecule is excited by polarized light, it emits light with a degree of polarization that is inversely proportional to the rate of molecular rotation. This property of fluorescence can be used to measure the interaction of a small labeled ligand with a larger protein and provides a basis for direct and competition binding assays. FP assays are readily adaptable to a high‐throughput format, have been used successfully in screens directed against a wide range of targets, and are particularly valuable in screening for inhibitors of protein‐protein and protein‐nucleic acid interactions when a small binding epitope can be identified for one of the partners. The protocols in this article describe a general procedure for development of FP assays to monitor binding of such a peptide or oligonucleotide to a protein of interest. Curr. Protoc. Chem Biol . 1:1‐15. © 2009 by John Wiley & Sons, Inc.