title: Monitoring dynamic binding of chromatin proteins in vivo by fluorescence recovery after photobleaching.
abstract: Fluorescence recovery after photobleaching (FRAP) has now become widely used to investigate nuclear protein binding to chromatin in live cells. FRAP can be applied qualitatively to assess if chromatin binding interactions are altered by various biological perturbations. It can also be applied semi-quantitatively to allow numerical comparisons between FRAP curves, and even fully quantitatively to yield estimates of in vivo diffusion constants and nuclear protein binding rates to chromatin. Here we describe how FRAP data should be collected and processed for these qualitative, semi-quantitative, and quantitative analyses.
authors.affiliation: Groupe Imagerie et Modélisation, Institut Pasteur, CNRS, URA 2582, Paris, France.
authors.forename: Florian
authors.initials: F
authors.lastname: Mueller
authors.forename: Tatiana S
authors.initials: TS
authors.lastname: Karpova
authors.forename: Davide
authors.initials: D
authors.lastname: Mazza
authors.forename: James G
authors.initials: JG
authors.lastname: McNally
mesh_terms: D002843:Chromatin / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D002843:Chromatin; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins