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PMID: 21284925 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

BioVyon Protein A, an alternative solid-phase affinity matrix for chromatin immunoprecipitation.

Analytical biochemistry ·Vol. 412 ·No. 2 ·2011-05-15 ·Pages 183-8

Chernukhin I, Kang SY, Brown S, Gretton S, Mendez-Catala CF, Cowieson D, Klenova E

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.

MeSH Terms
Animals Cell Line, Tumor Chromatin Immunoprecipitation/methods Chromatography, Affinity DNA/metabolism Humans Mice NIH 3T3 Cells Polyethylenes/metabolism Polymerase Chain Reaction Staphylococcal Protein A/metabolism
Chemicals
Polyethylenes Staphylococcal Protein A DNA
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Chernukhin Igor
Department of Biological Sciences, University of Essex, Colchester, Essex CO4 3SQ, UK.
Kang Sung Yun
Brown Sam
Gretton Svetlana
Mendez-Catala Claudia Fabiola
Cowieson Dave
Klenova Elena
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
1096-0309
Published
2011-05-15
Epub
2011-00-01
Pages
183-8
Language
English
Region
United States
NLM ID
0370535
Subset
IM
Grants
Cancer Research UK · 12723 · United Kingdom
Medical Research Council · G0401088 · United Kingdom
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