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Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin
Conventional chromatin immunoprecipitation (ChIP) includes many steps that need to be optimized. Here, we have described a protocol of fractionation-assisted native ChIP (fanChIP) which combines subfractionation and native ChIP to purify protein/chromatin complexes applicable for analyses of both pr...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8024768/ https://www.ncbi.nlm.nih.gov/pubmed/33855306 http://dx.doi.org/10.1016/j.xpro.2021.100404 |
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author | Miyamoto, Ryo Yokoyama, Akihiko |
author_facet | Miyamoto, Ryo Yokoyama, Akihiko |
author_sort | Miyamoto, Ryo |
collection | PubMed |
description | Conventional chromatin immunoprecipitation (ChIP) includes many steps that need to be optimized. Here, we have described a protocol of fractionation-assisted native ChIP (fanChIP) which combines subfractionation and native ChIP to purify protein/chromatin complexes applicable for analyses of both protein-protein and protein-DNA interactions within a short period of time. fanChIP is advantageous as subcellular fractionation removes chromatin-unbound materials before immunoprecipitation, and the chromatin fragmentation by micrococcal nuclease (MNase) in a mild condition enables one-step purification of intact protein/chromatin complexes. For complete details on the use and execution of this protocol, please refer to Miyamoto et al. (2020). |
format | Online Article Text |
id | pubmed-8024768 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-80247682021-04-13 Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin Miyamoto, Ryo Yokoyama, Akihiko STAR Protoc Protocol Conventional chromatin immunoprecipitation (ChIP) includes many steps that need to be optimized. Here, we have described a protocol of fractionation-assisted native ChIP (fanChIP) which combines subfractionation and native ChIP to purify protein/chromatin complexes applicable for analyses of both protein-protein and protein-DNA interactions within a short period of time. fanChIP is advantageous as subcellular fractionation removes chromatin-unbound materials before immunoprecipitation, and the chromatin fragmentation by micrococcal nuclease (MNase) in a mild condition enables one-step purification of intact protein/chromatin complexes. For complete details on the use and execution of this protocol, please refer to Miyamoto et al. (2020). Elsevier 2021-03-23 /pmc/articles/PMC8024768/ /pubmed/33855306 http://dx.doi.org/10.1016/j.xpro.2021.100404 Text en © 2021 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Miyamoto, Ryo Yokoyama, Akihiko Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin |
title | Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin |
title_full | Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin |
title_fullStr | Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin |
title_full_unstemmed | Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin |
title_short | Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin |
title_sort | protocol for fractionation-assisted native chip (fanchip) to capture protein-protein/dna interactions on chromatin |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8024768/ https://www.ncbi.nlm.nih.gov/pubmed/33855306 http://dx.doi.org/10.1016/j.xpro.2021.100404 |
work_keys_str_mv | AT miyamotoryo protocolforfractionationassistednativechipfanchiptocaptureproteinproteindnainteractionsonchromatin AT yokoyamaakihiko protocolforfractionationassistednativechipfanchiptocaptureproteinproteindnainteractionsonchromatin |