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完整的工作流程为组蛋白翻译后修饰的分析采用自下而上质谱法：从组蛋白提取数据分析
JoVE Journal
Biologia
This content is Free Access.
JoVE Journal Biologia
Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis
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完整的工作流程为组蛋白翻译后修饰的分析采用自下而上质谱法：从组蛋白提取数据分析

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

Capitoli

  • 00:05Titolo
  • 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

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这个协议概述了表征使用质谱（MS）组蛋白的翻译后修饰的完全集成的工作流程。该工作流包括使用纳米流液相色谱法和用于数据分析的指令的细胞培养物或组织中，组蛋白衍生和消化，MS分析组蛋白纯化。该协议被设计为内完成2 - 3天。

Tags

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

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