Mapping Nuclear Lamin-Genome Interactions by Chromatin Immunoprecipitation of Nuclear Lamins

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1411)

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.

Key words

Chromatin Chromatin immunoprecipitation Nuclear lamina Lamin A/C Lamin B Antibody Sonication 

Notes

Acknowledgments

Our work is supported by the Norwegian Research Council, the University of Oslo and the Norwegian Center for Stem Cell Research (P.C.), and by South East Health Norway (A.R.O.).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Molecular Medicine, Institute of Basic Medical SciencesUniversity of OsloOsloNorway

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