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© Research
Publication : Methods in molecular biology (Clifton, N.J.)

Monitoring dynamic binding of chromatin proteins in vivo by fluorescence recovery after photobleaching

Team: Imaging and Modeling
Team: Quantitative RNA imaging
Member: Florian Muller
Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Methods in molecular biology (Clifton, N.J.) - 01 Jan 2012

Mueller F, Karpova TS, Mazza D, McNally JG

Link to Pubmed [PMID] – 22183594

Methods Mol. Biol. 2012;833:153-76

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.