MOBE-ChIP: a large-scale chromatin immunoprecipitation assay for cell type-specific studies.
Cell type-specific transcriptional regulators play critical roles in the generation and maintenance of multicellularity. As they are often expressed at low levels, in vivo DNA-binding studies of these regulators by standard chromatin immunoprecipitation (ChIP) assays are technically challenging. We describe here an optimized ChIP protocol termed Maximized Objects for Better Enrichment (MOBE)-ChIP, which enhances the sensitivity of ChIP assays for detecting cell type-specific signals. The protocol, which is based on the disproportional increase of target signals over background at higher scales, uses substantially greater volume of starting materials than conventional ChIPs to achieve high signal enrichment. This technique can capture weak binding events that are ambiguous in standard ChIP assays, and is useful both in gene-specific and whole-genome analysis. This protocol has been optimized for Arabidopsis, but should be applicable to other model systems with minor modifications. The full procedure can be completed within 3 days.
Department of Biology, Stanford University, Stanford, CA, 94305, USA.
On Sun
http://orcid.org/0000-0002-8121-235X
OS
Lau
Department of Biology, Stanford University, Stanford, CA, 94305, USA.|Howard Hughes Medical Institute, Stanford University, Stanford, CA, 94305, USA.
Dominique C
DC
Bergmann
D017360:Arabidopsis / Q000378:metabolism; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D010940:Plant Proteins / Q000378:metabolism
D010940:Plant Proteins
Arabidopsis thailiana; ChIP-Seq; cell type-specific transcription factor/regulator; chromatin immunoprecipitation (ChIP) assay; technical advance