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

Chernukhin, Igor and Kang, Sung Yun and Brown, Sam and Gretton, Svetlana and Mendez-Catala, Claudia Fabiola and Cowieson, Dave and Klenova, Elena (2011) 'BioVyon Protein A, an alternative solid-phase affinity matrix for chromatin immunoprecipitation.' Analytical Biochemistry, 412 (2). pp. 183-188. ISSN 0003-2697

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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. © 2011 Published by Elsevier Inc. All rights reserved.

Item Type: Article
Uncontrolled Keywords: Chromatin immunoprecipitation (ChIP); BioVyon Protein A; Protein A Sepharose; Dynabeads Protein A; CCCTC-binding factor (CTCF); CTCF binding (target) sites (CTSs); RNA polymerase II; TATA box; Quantitative real-time polymerase chain reaction (Q-PCR)
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Science and Health
Faculty of Science and Health > Life Sciences, School of
SWORD Depositor: Elements
Depositing User: Elements
Date Deposited: 04 Aug 2011 15:06
Last Modified: 18 Aug 2022 11:01
URI: http://repository.essex.ac.uk/id/eprint/380

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