title: BioVyon Protein A, an alternative solid-phase affinity matrix for chromatin immunoprecipitation.
abstract: Chromatin immunoprecipitation (ChIP) is an important technique in the study of DNA/protein interactions. The ChIP procedure, however, has limitations in that it is lengthy, can be inconsistent, and is prone to nonspecific binding of DNA and proteins to the bead-based solid-phase matrices that are often used for the immunoprecipitation step. In this investigation, we examined the utility of a new matrix for ChIP assays, BioVyon Protein A, a solid support based on porous polyethylene. In ChIP experiments carried out using two antibodies and seven DNA loci, the performance of BioVyon Protein A was significantly better, with a greater percentage of DNA pull-down in all of the assays tested compared with bead-based matrices, Protein A Sepharose, and Dynabeads Protein A. Furthermore, the rigid porous disc format within a column made the BioVyon matrix much easier to use with fewer steps and less equipment requirements, resulting in a significant reduction in the time taken to process the ChIP samples. In summary, BioVyon Protein A provides a column-based assay method for ChIP and other immunoprecipitation-based procedures; the rigid porous structure of BioVyon enables a fast and robust protocol with higher ChIP enrichment ratios.
authors.affiliation: Department of Biological Sciences, University of Essex, Colchester, Essex CO4 3SQ, UK.
authors.forename: Igor
authors.initials: I
authors.lastname: Chernukhin
authors.forename: Sung Yun
authors.initials: SY
authors.lastname: Kang
authors.forename: Sam
authors.initials: S
authors.lastname: Brown
authors.forename: Svetlana
authors.initials: S
authors.lastname: Gretton
authors.forename: Claudia Fabiola
authors.initials: CF
authors.lastname: Mendez-Catala
authors.forename: Dave
authors.initials: D
authors.lastname: Cowieson
authors.forename: Elena
authors.initials: E
authors.lastname: Klenova
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D011095:Polyethylenes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D013205:Staphylococcal Protein A / Q000378:metabolism*
chemical_list: D011095:Polyethylenes; D013205:Staphylococcal Protein A; D004247:DNA