Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events.
The chromatin immunoprecipitation (ChIP) assay is a major tool in the study of genomic processes in vivo. This and other methods are revealing that control of gene expression, cell division and DNA repair involves multiple proteins and great number of their modifications. ChIP assay is traditionally done in test tubes limiting the ability to study signaling of the complex genomic events. To increase the throughput and to simplify the assay we have developed a microplate-based ChIP (Matrix ChIP) method, where all steps from immunoprecipitation to DNA purification are done in microplate wells without sample transfers. This platform has several important advantages over the tube-based assay including very simple sample handling, high throughput, improved sensitivity and reproducibility, and potential for automation. 96 ChIP measurements including PCR can be done by one researcher in one day. We illustrate the power of Matrix ChIP by parallel profiling 80 different chromatin and transcription time-course events along an inducible gene including transient recruitment of kinases.
Flanagin
Steve
S
UW Medicine Lake Union, Department of Medicine, University of Washington, Seattle, WA 98109, USA.
Nelson
Joel D
JD
Castner
David G
DG
Denisenko
Oleg
O
Bomsztyk
Karol
K
D000818:Animals; D000906:Antibodies / Q000737:chemistry; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D004247:DNA / Q000302:isolation & purification; D051766:Early Growth Response Protein 1 / Q000235:genetics; D007700:Kinetics; D011494:Protein Kinases / Q000032:analysis*; D051381:Rats; D015398:Signal Transduction; D014158:Transcription, Genetic
D000906:Antibodies; D002843:Chromatin; D051766:Early Growth Response Protein 1; D004247:DNA; D011494:Protein Kinases