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. DOI https://doi.org/10.1016/j.ab.2011.01.036
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. DOI https://doi.org/10.1016/j.ab.2011.01.036
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. DOI https://doi.org/10.1016/j.ab.2011.01.036
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: | Unnamed user with email elements@essex.ac.uk |
Depositing User: | Unnamed user with email elements@essex.ac.uk |
Date Deposited: | 04 Aug 2011 15:06 |
Last Modified: | 07 Aug 2024 19:42 |
URI: | http://repository.essex.ac.uk/id/eprint/380 |