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Methods. 2019 Oct 9. pii: S1046-2023(19)30136-7. doi: 10.1016/j.ymeth.2019.10.001. [Epub ahead of print]

Enrichment of histones from patient samples for mass spectrometry-based analysis of post-translational modifications.

Author information

1
Department of Experimental Oncology, IEO, European Institute of Oncology IRCCS, Milan, Italy. Electronic address: roberta.noberini@ieo.it.
2
Department of Experimental Oncology, IEO, European Institute of Oncology IRCCS, Milan, Italy. Electronic address: camilla.restellini@ieo.it.
3
Department of Experimental Oncology, IEO, European Institute of Oncology IRCCS, Milan, Italy. Electronic address: evelynoliva.savoia@ieo.it.
4
Department of Experimental Oncology, IEO, European Institute of Oncology IRCCS, Milan, Italy. Electronic address: tiziana.bonaldi@ieo.it.

Abstract

Aberrations in histone post-translational modifications (PTMs) have been implicated with the development of numerous pathologies, including cancer. Therefore, profiling histone PTMs in patient samples could provide information useful for the identification of epigenetic biomarkers, as well as for the discovery of potential novel targets. While antibody-based methods have been traditionally employed to analyze histone PTM in clinical samples, mass spectrometry (MS) can provide a more comprehensive, unbiased and quantitative view on histones and their PTMs. To combine the power of MS-based methods and the potential offered by histone PTM profiling of clinical samples, we have recently developed a series of methods for the extraction and enrichment of histones from different types of patient samples, including formalin-fixed paraffin-embedded tissues, fresh- and optimal cutting temperature-frozen tissues, and primary cells. Here, we provide a detailed description of these protocols, together with indications on the expected results and the most suitable workflow to be used downstream of each procedure.

KEYWORDS:

Epigenetics; Formalin-fixed paraffin-embedded tissues; Frozen tissues; Histone post-translational modifications; Mass spectrometry

PMID:
31605746
DOI:
10.1016/j.ymeth.2019.10.001
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