Mapping chromosomal proteins in vivo by formaldehyde-crosslinked-chromatin immunoprecipitation.
Gene regulation is a complex process. Numerous factors appear to be required for the accurate temporal and spatial regulation of each gene. Often these factors are assembled into multiprotein complexes, contributing to specific gene regulation events. Understanding how all these factors are organized in the chromosome and how their function is regulated in vivo is a challenging task. One of the most useful techniques for studying this level of gene regulation is the in vivo fixation by formaldehyde crosslinking of proteins to proteins and proteins to DNA, followed by immunoprecipitation of the fixed material.
Orlando
V
V
DIBIT HSR Biomedical Scientific Park, Via Olgettina 58, 20132 Milano, Italy. v.orlando@hsr.it
D000818:Animals; D015139:Blotting, Southern; D002499:Centrifugation, Density Gradient / Q000379:methods; D002586:Cesium; D002712:Chlorides; D002843:Chromatin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis* / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry; D005786:Gene Expression Regulation; D010449:Peptide Mapping / Q000379:methods*; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests / Q000379:methods*
D002712:Chlorides; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D003432:Cross-Linking Reagents; D005557:Formaldehyde; D002586:Cesium; C028019:cesium chloride