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Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis
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Biology
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Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis

DOI:
10.3791/54112-v

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11:02 min

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May 17, 2016

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, , , ,
1Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine,University of Pennsylvania

Chapters

  • 00:05Title
  • 01:07Isolation of Nuclei from Intact Cells
  • 03:18Extraction and Purification of Histones from Nuclei
  • 05:53Chemical Derivatization of Histones Using Propionic Anhydride for Bottom-up Analysis
  • 07:43Sample Desalting with Stage-tips
  • 08:59Results: Characterization of Histone Post-translational Modifications
  • 10:10Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol outlines a fully integrated workflow for characterizing histone post-translational modifications using mass spectrometry (MS). The workflow includes histone purification from cell cultures or tissues, histone derivatization and digestion, MS analysis using nano-flow liquid chromatography and instructions for data analysis. The protocol is designed for completion within 2 - 3 days.

Tags

Keywords: HistonePost-translational ModificationsMass SpectrometryBottom-up ProteomicsHistone ExtractionChromatin BiologyGene Expression RegulationChromosome CondensationEpigenetic ChangesChromatin OperationCell CultureHistone DerivatizationHistone DigestionNanoflow Liquid Chromatography

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