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

Anal Biochem. 2011 May 15;412(2):183-8. doi: 10.1016/j.ab.2011.01.036. Epub 2011 Feb 1.

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.

Publication types

  • Research Support, Non-U.S. Gov't

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*

Substances

  • Polyethylenes
  • Staphylococcal Protein A
  • DNA