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Specific, sensitive and quantitative protein detection by in-gel fluorescence

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  • Adrian C. D. Fuchs

    (Max Planck Institute for Biology)

Abstract

Recombinant proteins in complex solutions are typically detected with tag-specific antibodies in Western blots. Here we describe an antibody-free alternative in which tagged proteins are detected directly in polyacrylamide gels. For this, the highly specific protein ligase Connectase is used to selectively fuse fluorophores to target proteins carrying a recognition sequence, the CnTag. Compared to Western blots, this procedure is faster, more sensitive, offers a better signal-to-noise ratio, requires no optimization for different samples, allows more reproducible and accurate quantifications, and uses freely available reagents. With these advantages, this method represents a promising alternative to the state of the art and may facilitate studies on recombinant proteins.

Suggested Citation

  • Adrian C. D. Fuchs, 2023. "Specific, sensitive and quantitative protein detection by in-gel fluorescence," Nature Communications, Nature, vol. 14(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-38147-8
    DOI: 10.1038/s41467-023-38147-8
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    References listed on IDEAS

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    1. Monya Baker, 2015. "Reproducibility crisis: Blame it on the antibodies," Nature, Nature, vol. 521(7552), pages 274-276, May.
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