title: Improving chromatin immunoprecipitation (ChIP) by suppression of method-induced DNA-damage signaling.
abstract: Genomic DNA is always associated with proteins that modulate the accessibility of the genetic information. This chromatin is the essential structure in which all nuclear activity from regulation to replication, transcription, and repair takes place. This dynamic structure can be most efficiently analyzed by using the method of chromatin immunoprecipitation (ChIP), where application of cell-permeable cross-linkers to living cells induces covalent bridging between proteins and adjacent DNA in the nucleus. After fragmentation of the DNA, the complexed proteins are isolated by binding to specific antibodies. The attached DNA is isolated and can be analyzed. This method has been improved multiple times and adjusted to different experimental needs. This chapter describes a further advance based on the observation that the current standard method itself induces alterations in the chromatin.
authors.affiliation: Institute of Veterinary Pharmacology and Toxicology/Vetsuisse, University of Zurich, Winterthurerstrasse 260, CH-8057, Zurich, Switzerland, sascha.beneke@vetpharm.uzh.ch.
authors.forename: Sascha
authors.initials: S
authors.lastname: Beneke
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D002725:Chloroform / Q000737:chemistry; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004249:DNA Damage; D053938:DNA Fragmentation; D006801:Humans; D019800:Phenol / Q000737:chemistry; D016133:Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D019800:Phenol; D002725:Chloroform; D004247:DNA