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

Adrian C. D. Fuchs ()
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Adrian C. D. Fuchs: Max Planck Institute for Biology

Nature Communications, 2023, vol. 14, issue 1, 1-10

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-38147-8

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