Method | Utility | Benefits | Drawbacks |
---|---|---|---|
Fluorescence recovery after photobleaching (FRAP) | Measurement of chromatin protein binding kinetics | 1. Can be used for nucleosomes as well as other chromatin binding proteins 2. Allows observation of protein location within the nucleus | 1. Cannot determine the specific genomic sites that are bound 2. Requires an epitope-tagged protein that may not behave exactly like the native form |
MS-based kinetic methods | Measurements of histone modification kinetics | Can be used for nucleosomes as well as other chromatin-binding proteins | Cannot determine the kinetics at specific genomic sites |
Inducible transgene-based methods | Measurement of nucleosome turnover kinetics as well as binding of other chromatin proteins | Can be used for nucleosomes as well as other chromatin-binding proteins | 1. Requires an epitope-tagged protein 2. Time lag during induction limits time resolution |
Recombination-induced tag exchange (RITE) | Measurement of nucleosome turnover kinetics as well as binding of other chromatin proteins | Can be used for nucleosomes as well as other chromatin-binding proteins | 1. Requires an epitope-tagged protein that may not behave exactly like the native form 2. Time lag during recombination limits time resolution |
Covalent attachment of tags to capture histones and identify turnover (CATCH-IT) | Measurement of nucleosome turnover kinetics | 1. No transgenes or antibodies are required 2. Excellent time resolution 3. Can be used on many different cell types | Only (H3/H4)2 tetramer incorporation kinetics can be measured easily |