[Abstract]  [Full Text]  [PDF]

Display
[Abstract]
[Full Text]
PDF
Search in Pubmed
Author: Ricke, R
Author: Bielinsky, A
Related works

Methods Article

Easy detection of chromatin binding proteins by the histone association assay

Robin Ricke1 and Anja-Katrin Bielinsky1*

1 Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota. Minneapolis, MN 55455. USA.

* To whom correspondence should be addressed: Anja-Katrin Bielinsky, Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota. 321 Church Street SE, 6-155 Jackson Hall, Minneapolis, MN 55455. USA. Phone: 612-624-2469. Fax: 612-625-2163. Email: bielinsk@cbs.umn.edu

Biol. Proced. Online 2005;7:60-69. doi:10.1251/bpo106
Submitted: March 22, 2005; Accepted: April 14, 2005; Published: May 09, 2005.

Indexing terms: Chromatin; Cross-linking Reagents; Formaldehyde; Histones; Immunoprecipitation.

Abbreviations: ChIP, Chromatin Immunoprecipitation; HAA, Histone Association Assay; Mcm10, Minichromosome Maintenance Protein 10; Orc2, Origin Recognition Complex protein 2; WCE, Whole Cell Extract.


Figure 2 Enlarged

Fig. 2:

Extracts are efficiently crosslinked with formaldehyde after 15 minutes of treatment. ABy025 cells (MCM10-9MYC) were arrested in G1 phase of the cell cycle by the addition of alpha factor. Mcm10-9Myc and histone H2B were detected by Western blotting in extracts quenched with glycine after 15 minutes or 24 hours of formaldehyde crosslinking. Crosslinks in WCEs (Input) and histone H3-immunoprecipitates (H3 IP) were reversed prior to SDS-PAGE. Myc (Labvision Neomarkers) and histone H2B (Upstate) specific antibodies were used for immunoblotting, respectively. Half of the input controls and all of the histone H3 immunoprecipitates were loaded.

Disclaimer/Legal Copyright Privacy Policy

ISSN: 1480-9222
Article © by the author(s). This paper is Open Access and is published in Biological Procedures Online under license from the author(s). Copying, printing, redistribution and storage permitted. Journal © 1997-2007 Biological Procedures Online.