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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...

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Detalles Bibliográficos
Autores principales: Miyamoto, Ryo, Yokoyama, Akihiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
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).
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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
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