Title: Sequential Chromatin Immunoprecipitation from Animal Tissues
Abstract: This chapter presents two different protocols for formaldehyde crosslinking of mouse tissue chromatin, followed by methods for chromatin solubilization, purification, fragmentation, and immunoprecipitation; some of these steps require special adaptations for animal tissues. The chromatin immunoprecipitation (ChIP) technique for identifying transcription factors and histones bound to specific DNA sites in cells has revolutionized the analysis of gene regulation. In this procedure, cells or nuclei are exposed to formaldehyde, which crosslinks proteins bound to each other and to DNA in native chromatin. The crosslinked chromatin is isolated and fractionated, the proteins are immunoprecipitated and assessed by a quantitative polymerase chain reaction (PCR) to determine the relative extent to which the antigen is associated with the DNA. Although the methods presented in the chapter were developed for liver chromatin, these methods represent a good starting point for working with other tissue types. Despite the technical obstacles regarding the handling of animal tissues for ChIP assays, a growing number of laboratories have made advances in the area.
Publication Year: 2003
Publication Date: 2003-01-01
Language: en
Type: review
Indexed In: ['crossref', 'pubmed']
Access and Citation
Cited By Count: 38
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