Species: Hu, Mu, Ba
Applications: IP, ChIP
Description
Chromatin Immunoprecipitation (ChIP) is a powerful method used to identify regions of the genome associated with specific proteins. These associations are crucial for vital cellular functions including gene transcription, DNA replication and recombination, repair, segregation, chromosomal stability, cell cycle progression, and epigenetic silencing. The ChIP assay has been used to study both histones and non-histone proteins, such as transcription factors, within the context of the cell. Transcription factors and other DNA binding proteins have a weaker affinity than histones, which generally are tightly associated within the chromatin complex. To avoid dissociation of non-histone proteins from the chromatin binding site, it is necessary to incorporate a cross-link step. QuikChIP(TM) Kit has been optimized for use with both histone and non-histone proteins. Using the QuikChIP(TM) Assay, cells are fixed with formaldehyde to cross-link nonhistone proteins to DNA. Alternatively, the cross-linking steps are not needed for histone proteins, which are tightly bound to DNA. The chromatin is released from the nuclei and sheared into 200-1000 bp fragments. The chromatin is then immunoprecipitated with specific antibodies. DNA sequences cross-linked to the protein of interest co-precipitate as part of the chromatin complex. The DNA/Chromatin/Antibody complex is then isolated using Protein G Agarose. After reverse cross-linking, the associated DNA is released from the complex and is ready for analysis.
