Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Blotting protein complexes from native gels to electron microscopy grids

Nature Methods volume 9, pages 182–184 (2012)Cite this article

Subjects

Abstract

We report a simple and generic method for the direct transfer of protein complexes separated by native gel electrophoresis to electron microscopy grids. After transfer, sufficient material remains in the gel for identification and characterization by mass spectrometry. The method should facilitate higher-throughput single-particle analysis by substantially reducing the time needed for protein purification, as demonstrated for three complexes from Thermoplasma acidophilum.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Grid-blotting procedure.
Figure 2: Electron micrographs of protein complexes blotted from native gels.

Similar content being viewed by others

References

  1. Nickell, S. et al. FEBS Lett. 581, 2751–2756 (2007).

    Article  CAS  Google Scholar 

  2. Frank, J. Q. Rev. Biophys. 42, 139–158 (2009).

    Article  CAS  Google Scholar 

  3. Henderson, R. Q. Rev. Biophys. 28, 171–193 (1995).

    Article  CAS  Google Scholar 

  4. Carragher, B. et al. J. Struct. Biol. 132, 33–45 (2000).

    Article  CAS  Google Scholar 

  5. Schagger, H., Cramer, W.A. & Vonjagow, G. Anal. Biochem. 217, 220–230 (1994).

    Article  CAS  Google Scholar 

  6. Sun, N. et al. Proteomics 9, 3783–3786 (2009).

    Article  CAS  Google Scholar 

  7. Phipps, B.M., Hoffmann, A., Stetter, K.O. & Baumeister, W. EMBO J. 10, 1711–1722 (1991).

    Article  CAS  Google Scholar 

  8. Dahlmann, B. et al. FEBS Lett. 251, 125–131 (1989).

    Article  CAS  Google Scholar 

  9. Pamnani, V. et al. FEBS Lett. 404, 263–268 (1997).

    Article  CAS  Google Scholar 

  10. Taylor, K.A. & Glaeser, R.M. J. Struct. Biol. 163, 214–223 (2008).

    Article  CAS  Google Scholar 

  11. Christiansen, C., Freundt, E.A. & Black, F.T. Int. J. Syst. Bacteriol. 25, 99–101 (1975).

    Article  Google Scholar 

  12. Nickell, S. et al. J. Struct. Biol. 149, 227–234 (2005).

    Article  Google Scholar 

Download references

Acknowledgements

We acknowledge the support of G. Pfeifer for data acquisition at the Titan Krios electron microscope. This work was supported in part by funding from the EU 7th Framework Program Proteomics Specification in Space and Time grant (PROSPECTS HEALTH-F4-2008-201648).

Author information

Author notes

  1. Roland Wilhelm Knispel, Christine Kofler & Stephan Nickell

    Present address: Present addresses: ChemAxon Kft., Budapest, Hungary (R.W.K.), Tietz Video and Image Processing Systems GmbH, Gauting, Germany (C.K.) and Carl Zeiss NTS GmbH, Oberkochen, Germany (S.N.).,

Authors and Affiliations

  1. Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany

    Roland Wilhelm Knispel, Christine Kofler, Marius Boicu, Wolfgang Baumeister & Stephan Nickell

Authors
  1. Roland Wilhelm Knispel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christine Kofler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marius Boicu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wolfgang Baumeister
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stephan Nickell
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

R.W.K., C.K. and S.N. designed and conducted the grid-blotting experiments. R.W.K. and M.B. prepared protein samples. R.W.K. and S.N. acquired electron microscopy data, analyzed images and prepared the figures. R.W.K., W.B. and S.N. wrote the manuscript.

Corresponding author

Correspondence to Wolfgang Baumeister.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 and Supplementary Table 1 (PDF 1752 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Knispel, R., Kofler, C., Boicu, M. et al. Blotting protein complexes from native gels to electron microscopy grids. Nat Methods 9, 182–184 (2012). https://doi.org/10.1038/nmeth.1840

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1840

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing