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Anal Biochem ; 412(2): 183-8, 2011 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-21284925

RESUMO

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.


Assuntos
Imunoprecipitação da Cromatina/métodos , Cromatografia de Afinidade , Polietilenos/metabolismo , Proteína Estafilocócica A/metabolismo , Animais , Linhagem Celular Tumoral , DNA/metabolismo , Humanos , Camundongos , Células NIH 3T3 , Reação em Cadeia da Polimerase
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