title: Dual cross-linking ribonucleoprotein immunoprecipitation assay.
abstract: Ribonucleoprotein immunoprecipitation (RIP) is an antibody-based method to detect RNA-protein interactions in situ. In the assay, UV cross-linking is commonly used to preserve RNA-protein interactions for subsequent target identification. UV light is a zero-length cross linker and thus identifies proteins directly bound to RNAs. Here, we describe a dual cross-linking RIP method that involves sequential protein-protein cross-linking step with a protein-protein cross-linker, followed by protein-RNA fixation by UV irradiation. In this way, proteins that indirectly bound to RNA can be analyzed.
authors.affiliation: Manitoba Institute of Child Health, 715 McDermot Avenue, University of Manitoba, Winnipeg, MB R3E 3P4, Canada.
authors.forename: Dilshad H
authors.initials: DH
authors.lastname: Khan
authors.forename: James R
authors.initials: JR
authors.lastname: Davie
mesh_terms: D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000737:chemistry; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D012313:RNA / Q000302:isolation & purification; D012261:Ribonucleoproteins / Q000737:chemistry / Q000302:isolation & purification*; D013388:Succinimides / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D012261:Ribonucleoproteins; D013388:Succinimides; D012313:RNA; C011240:dithiobis(succinimidylpropionate)
keywords: RNA-protein interaction; UV cross-linking; double pontage; dual cross-linking; immunoprecipitation; immunoprécipitation; interaction ARN-protéine; pontage par UV; ribonucleoprotein; ribonucléoprotéine