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Tricine–SDS-PAGE

Nature Protocols volume 1, pages 16–22 (2006)Cite this article

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Abstract

Tricine–SDS-PAGE is commonly used to separate proteins in the mass range 1–100 kDa. It is the preferred electrophoretic system for the resolution of proteins smaller than 30 kDa. The concentrations of acrylamide used in the gels are lower than in other electrophoretic systems. These lower concentrations facilitate electroblotting, which is particularly crucial for hydrophobic proteins. Tricine–SDS-PAGE is also used preferentially for doubled SDS-PAGE (dSDS-PAGE), a proteomic tool used to isolate extremely hydrophobic proteins for mass spectrometric identification, and it offers advantages for resolution of the second dimension after blue-native PAGE (BN-PAGE) and clear-native PAGE (CN-PAGE). Here I describe a protocol for Tricine–SDS-PAGE, which includes efficient methods for Coomassie blue or silver staining and electroblotting, thereby increasing the versatility of the approach. This protocol can be completed in 1–2 d.

*Note: In the version of the article initially published online, the words “Gel buffer (3x)” were missing in the table on page 18.  The error has been corrected in all versions of the article.

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Figure 1: Comparison of Tricine–SDS-PAGE and Laemmli–SDS-PAGE.
Figure 2: Tricine–SDS-PAGE of marker proteins using various types of gel.

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Change history

  • 10 August 2006

    In the version of the article initially published online, the words “Gel buffer (3x)” were missing in the table on page 18. The error has been corrected in all versions of the article.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 628, Project P13.

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  1. Molekulare Bioenergetik, Zentrum der Biologischen Chemie, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, Haus 26, Frankfurt, D-60590, Germany

    Hermann Schägger

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Correspondence to Hermann Schägger.

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Schägger, H. Tricine–SDS-PAGE. Nat Protoc 1, 16–22 (2006). https://doi.org/10.1038/nprot.2006.4

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