Abstract
Purification of small, native chromatin regions for proteomic identification of specifically bound proteins and histone posttranslational modifications is a powerful approach for studying mechanisms of chromosome metabolism. Here we detail a Chromatin Affinity Purification with Mass Spectrometry (ChAP-MS) approach for affinity purification of 1 kb regions of chromatin for targeted proteomic analysis. This approach utilizes quantitative, high resolution mass spectrometry to categorize proteins and histone posttranslational modifications co-enriched with the given chromatin region as either “specific” to the targeted chromatin or “nonspecific” contamination. In this way, the ChAP-MS approach can help define and redefine mechanisms of chromatin-templated activities.
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Acknowledgments
We would like to acknowledge support for mass spectrometry from the UAMS Proteomics Facility and support from NIH grants R01GM106024, R33CA173264, UL1RR029884, P30GM103450, and P20GM103429.
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Byrum, S.D., Taverna, S.D., Tackett, A.J. (2015). Purification of Specific Chromatin Loci for Proteomic Analysis. In: Hancock, R. (eds) The Nucleus. Methods in Molecular Biology, vol 1228. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1680-1_8
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DOI: https://doi.org/10.1007/978-1-4939-1680-1_8
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-1680-1
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