Overview: Chromatin immunoprecipitation (ChIP) is a powerful tool for identifying proteins, including histone proteins and non-histone proteins, associated with specific regions of the genome by using specific antibodies that recognize a specific protein or a specific modification of a protein. The initial step of ChIP is the cross-linking of protein-protein and protein-DNA in live cells with formaldehyde. After cross-linking, the cells are lysed and crude extracts are sonicated to shear the DNA. Proteins together with cross-linked DNA are subsequently immunoprecipitated. Protein-DNA cross-links in the immunoprecipitated material are then reversed and the DNA fragments are purified and PCR amplified. There are two general types of ChIP experiments: Cross-linking ChIP (X-ChIP or XChIP) which uses chromatin fixed with formaldehyde and fragmented by sonication and native chromatin ChIP (N-ChIP or NChIP) which uses native chromatin prepared by nuclease digestion of cell nuclei.
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Protocols
- ChIP Assay (Farnham Laboratory, UC Davis)
This protocol has successfully used with NIH 3T3, Friend, HeLa, Raji and CHO cells. However, you may have to optimize conditions for your specific cell type.
http://genomecenter.ucdavis.edu/farnham/farnham/pr...
Added: Mon Mar 29 2004, Hits: 19263, Reviews: 0 Write review Cached - ChIP Assay Protocol (The Epigenome Network of Excellence)
Well-written chromatin immunoprecipitation protocol for histone modification & chromatin associated proteins.
http://www.epigenome-noe.net/researchtools/protoco...
Added: Sat Mar 18 2006, Hits: 3902, Reviews: 0 Write review Cached - ChIP Protocol
(UHN Microarray Center)
Well written and detailed ChIP and microarray protocol.
http://www.microarrays.ca/support/PDFs/ChIP_Protoc...
Added: Thu Jan 27 2005, Hits: 12695, Reviews: 0 Write review - Formaldehyde Crosslinking and ChIP Assay (Mirmira Laboratory at the University of Virginia)
If you want to start a ChIP experiment without buying a kit, here is the protocol to follow. It guides you through a 50-step protocol from formaldehyde crosslinking to ChIP and also provides receipes for all home-made reagents.
http://faculty.virginia.edu/mirmira/resources_file...
Added: Sat Jun 05 2004, Hits: 11665, Reviews: 0 Write review Cached - Standard ChIP Protocol with Modifications (The Epigenome Network of Excellence )
This is a general protocol for ChIP, also includes modifications such as re-ChIP, alternative crosslinking, andnNucleosome ChIPs.
http://www.epigenome-noe.net/researchtools/protoco...
Added: Sat Mar 18 2006, Hits: 2599, Reviews: 0 Write review Cached -
ChIP Protocol for Beginners
(Sugden Laboratory, McArdle Laboratory for Cancer Research)
new
http://mcardle.oncology.wisc.edu/sugden/protocols/...
Added: Sat Mar 01 2008, Hits: 42, Reviews: 0 Write review -
Chromatin Immunoprecipitation (ChIP) Assay; Quantitation of DNA
(Millipore)
new
Method of quantitating immunoprecipitated DNA by running acrylamide gels and stained with SYBR Green I
http://www.millipore.com/techpublications/tech1/mc...
Added: Sat Mar 01 2008, Hits: 11, Reviews: 0 Write review Cached -
MicroChIP Protocol
(Dr. Peggy Farnham, UC Davis Genome Center)
new
ChIP protocol for 1x10^4 to 1x10^5 cells.
http://genomecenter.ucdavis.edu/expression_analysi...
Added: Sat Mar 01 2008, Hits: 11, Reviews: 0 Write review -
ChIP
(OpenWetWare)
Chromatin Immunoprecipitation determines the in vivo chromatin binding sites of a transcription factor or other protein of interest.
http://openwetware.org/wiki/ChIP
Added: Sun Mar 04 2007, Hits: 1143, Reviews: 0 Write review Cached -
ChIP Assay
(The Morimoto Lab, Northwestern University)
http://www.biochem.northwestern.edu/ibis/morimoto/...
Added: Mon Mar 29 2004, Hits: 7965, Reviews: 0 Write review
Troubleshootings and FAQs
- ChIp assay on myocytes or tissues?
- Advice on ChIP assay
- Sonication settings for ChIP assay
- How to sonicate DNA in a water bath for ChIP
- ChIP sonication woes- Losing DNA?
- ChIP against transcription factors
- PCR for ChIP not working
- ChIP - Foaming in sonication and other woes
- Is proteinase K digestion necessary and what alternative to sonication for DNA shearing?
- ChIP DNA purification methods
- ChIP - no immunoprecipitated DNA
- MNase vs sonication for ChIP