Advancing Epigenetics Towards Systems Biology

Chromatin Immunoprecipitation (ChIP) Assay (Prot 11)

Antigone Kouskouti & Irene Kyrmizi

Introduction

Chromatin Immuno precipitation (ChIP) assays are used to evaluate the association of proteins with specific DNA regions. The technique involves crosslinking of proteins with DNA, fragmentation and preparation of soluble chromatin followed by immunoprecipitation with an antibody recognizing the protein of interest. The segment of the genome associated with the protein is then identified by PCR amplification of the DNA in the immunoprecipitates. There is a general protocol for ChIP, which is employed by many different labs with minor modifications. Here we describe modified ChIP protocols used successfully in our lab. They include the Standard Protocol, Re-ChIP, Alternative Crosslinking Method and Nucleosome ChIPs.

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Antigone Kouskouti & Irene Kyrmizi

Talianidis Lab - Institute of Molecular Biology and Biotechnology - FORTH - 1527 Vassilika Vouton - 711 10, Heraklion, Crete, Greece

Antigone Kouskouti & Irene Kyrmizi