Individual and Sequential Chromatin Immunoprecipitation Protocols
Por:
Furlan-Magaril M., Recillas-Targa F.
Publicada:
1 ene 2015
Resumen:
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 profi le 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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