title: Mapping Nuclear Lamin-Genome Interactions by Chromatin Immunoprecipitation of Nuclear Lamins.
abstract: The nuclear lamina is a meshwork of A- and B-type lamins which interact with chromatin and regulate many nuclear functions. Recent studies have reported the discovery of chromatin domains interacting with nuclear lamins by chromatin immunoprecipitation (ChIP) of lamin A/C or B1 and identification of the associated DNA sequences by microarray or high-throughput sequencing. ChIP has been used over many years to get a snapshot of interactions between DNA and proteins in cells, including modified histones, transcription factors, chromatin remodelers, and recently, structural proteins such as nuclear lamins. We describe here the procedure we have established in our laboratory for ChIP of lamin A/C and lamin B1 from human cultured cells. The protocol is compatible with polymerase chain reaction and high-throughput sequencing analysis of the co-immunoprecipitated DNA.
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Blindern, 0317, 1112, Oslo, Norway. a.r.oldenburg@medisin.uio.no.
authors.forename: Anja R
authors.initials: AR
authors.lastname: Oldenburg
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Blindern, 0317, 1112, Oslo, Norway. philippe.collas@medisin.uio.no.
authors.forename: Philippe
authors.initials: P
authors.lastname: Collas
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D016678:Genome; D059014:High-Throughput Nucleotide Sequencing; D034882:Lamins / Q000378:metabolism*; D034881:Nuclear Lamina / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D011485:Protein Binding
chemical_list: D002843:Chromatin; D034882:Lamins; D009687:Nuclear Proteins
keywords: Antibody; Chromatin; Chromatin immunoprecipitation; Lamin A/C; Lamin B; Nuclear lamina; Sonication