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Separation of the Plant Golgi Apparatus and Endoplasmic Reticulum by Free-Flow Electrophoresis

  • Harriet T. Parsons
  • Susana M. González Fernández-Niño
  • Joshua L. Heazlewood
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1072)

Abstract

Free-flow electrophoresis (FFE) is a technique for separation of proteins, peptides, organelles, and cells. With zone electrophoresis (ZE-FFE), organelles are separated according to surface charge. The plant Golgi and endoplasmic reticulum (ER) are similar in density and are therefore separated with difficulty using standard techniques such as density centrifugation. Purification of the ER and Golgi apparatus permits a biochemical and proteomic characterization which can reveal the division of processes between these compartments. Here we describe complete separation between the ER and more negatively charged Golgi compartments using ZE-FFE. We also describe techniques for assigning proteins to partially separated ER and the less negatively charged Golgi compartments.

Key words

Free-flow electrophoresis Golgi apparatus Arabidopsis Endoplasmic reticulum Proteomics 

Notes

Acknowledgments

This work conducted by the Joint BioEnergy Institute was supported by the Office of Science, Office of Biological and Environmental Research, of the US Department of Energy under Contract No. DE-AC02-05CH11231.

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Harriet T. Parsons
    • 1
  • Susana M. González Fernández-Niño
    • 1
  • Joshua L. Heazlewood
    • 1
  1. 1.Joint BioEnergy Institute and Physical Biosciences DivisionLawrence Berkeley National LaboratoryBerkeleyUSA

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