Separation and Purification of Multiply Acetylated Proteins Using Cation-Exchange Chromatography

  • Romeo Papazyan
  • Sean D. Taverna
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 981)

Abstract

High-performance liquid chromatography (HPLC) is extremely useful for the study of proteins and the characterization of their posttranslational modifications. Here we describe a method that utilizes cation-exchange HPLC to separate multiply acetylated histone H3 species on the basis of their charge and hydrophilicity. This high-resolution method allows for the separation of histone H3 species that differ by as few as one acetyl group, and is compatible with subsequent analysis by a variety of techniques, including mass spectrometry and western blotting.

Key words

Cation-exchange chromatography PolyCAT A Acetylation HPLC Histone H3 Acetic anhydride 

Notes

Acknowledgements

We thank T. Gilbert and other members of the Taverna laboratory for comments on this protocol. This work was supported by NIH grant RO190035489.

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Romeo Papazyan
    • 1
  • Sean D. Taverna
    • 1
  1. 1.Department of Pharmacology and Molecular SciencesJohns Hopkins University School of MedicineBaltimoreUSA

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