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Author: Ricke, R
Author: Bielinsky, A
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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 4 Enlarged

Fig. 4:

Using the histone association assay under quantitative conditions. A) Flowchart for the quantitative assessment of DNA bound protein(s). Crosslinked WCEs are titrated and immunoprecipitated with histone H3-specific antibodies. Both the supernatant (blue) and histone H3 immunoprecipitate (IP) (yellow) are monitored for DNA as well as histones H2B and H3. B) Bar graph representing the amounts of Orc2, histone H2B, histone H3 and Mcm10 in the input (maroon) and histone H3 immunoprecipitate (IP) (yellow) under quantitative conditions, as described (21).

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