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A Multichannel Gel Electrophoresis and Continuous Fraction Collection Apparatus for High-Throughput Protein Separation and Characterization

  • Ming Dong
  • Megan Choi
  • Mark D. Biggin
  • Jian Jin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 869)

Abstract

We developed a multichannel gel electrophoresis system that continuously collects fractions as protein bands migrate off the bottom of gel columns. The device uses several short linear gel columns, each of a different percent acrylamide, to achieve a separation power similar to that of a long gradient gel. A “Counter Free-Flow” elution technique allows continuous and simultaneous fraction collection from multiple channels at low cost. Using the system with SDS-PAGE, 300 μg samples of protein can be separated and eluted into 48–96 fractions over a mass range of 10–150 kDa in 2.5 h. Each eluted protein can be recovered at 50% efficiency or higher in ∼500 μL. The system can also be used for native gel electrophoresis, but protein aggregation limits the loading capacity to about 50 μg per channel and reduces resolution. This system has the potential to be coupled with mass spectrometry to achieve high-throughput protein identification.

Key words

Continuous elution electrophoresis Counter free-flow High-throughput Multichannel electrophoresis 

Notes

Acknowledgments

This work was part of ENIGMA, a Scientific Focus Area Program supported by the US Department of Energy, Office of Science, Office of Biological and Environmental Research, Genomics: GTL Foundational Science through contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the US Department of Energy.

References

  1. 1.
    Butland G, Peregrín-Alvarez JM, Li J et al (2005) Interaction network containing conserved and essential protein complexes in Escherichia coli. Nature 433:531–537PubMedCrossRefGoogle Scholar
  2. 2.
    Choi M, Nordmeyer R, Cornell E et al (2010) A multichannel gel electrophoresis and continuous fraction collection apparatus for high-throughput protein separation and characterization. Electro­phoresis 31:440–447PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Ming Dong
    • 1
  • Megan Choi
    • 1
  • Mark D. Biggin
    • 1
  • Jian Jin
    • 1
  1. 1.Lawrence Berkeley National LaboratoryBerkeleyUSA

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