Abstract
DNA regulatory elements nucleate the interaction of several transcription factors in conjunction with ubiquitous and/or tissue-specific cofactors in order to regulate gene expression making it relevant to determine the profiles of cohabitation of several proteins on the chromatin fiber. Chromatin immunoprecipitation (ChIP) has been broadly used to determine the profile of several histone posttranslational modifications as well as transcription factor occupancy in vivo. However, individual ChIP does not resolve whether the epitope under study is present at the same time on a given genomic location. Here we describe the ChIP-re-ChIP assay that represents a direct strategy to determine the in vivo co-localization of proteins or histone posttranslational modifications in a chromatinized template on the basis of double and independent rounds of immunoprecipitation with high-quality ChIP-grade antibodies.
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Acknowledgments
We would like to thank Georgina Guerrero and Fernanda Suaste Olmos for her excellent technical assistance. This work was supported by grants from the Dirección General de Asuntos del Personal Académico-UNAM (IN203811 and IN128464), Consejo Nacional de Ciencia y Tecnología, CONACyT (128464 and 220503).
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Furlan-Magaril, M., Recillas-Targa, F. (2015). Individual and Sequential Chromatin Immunoprecipitation Protocols. In: Leblanc, B., Rodrigue, S. (eds) DNA-Protein Interactions. Methods in Molecular Biology, vol 1334. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2877-4_13
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DOI: https://doi.org/10.1007/978-1-4939-2877-4_13
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2876-7
Online ISBN: 978-1-4939-2877-4
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