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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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Biología
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JoVE Revista Biología
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

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11:02 min

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May 17, 2016

DOI:

10.3791/54112-v

DOI:
10.3791/54112-v

•

11:02 min
May 17, 2016

•

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

Summary

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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.

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

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Sidoli, S., Bhanu, N. V., Karch, K. R., Wang, X., Garcia, B. A. Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis. J. Vis. Exp. (111), e54112, doi:10.3791/54112 (2016).

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