ArticleScientific reports2019
Quantification of very low-abundant proteins in bacteria using the HaloTag and epi-fluorescence microscopy.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 47 citations in OpenAlex.
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- In vivo single-molecule imaging of RecB reveals efficient repair of DNA damage in Escherichia coli.Nucleic acids research · 2025Article
- Experimental Approaches to Visualize Effector Protein Translocation During Host-Pathogen Interactions.BioEssays : news and reviews in molecular, cellular and developmental biology · 2025Review
- Real-time imaging of axonal membrane protein life cycles.Nature protocols · 2024Review
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- Double-Labeling Method for Visualization and Quantification of Membrane-Associated Proteins inInternational journal of molecular sciences · 2023Article
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- RecA and RecB: probing complexes of DNA repair proteins with mitomycin C in liveJournal of the Royal Society, Interface · 2022Article
- Quantifying biochemical reaction rates from static population variability within incompletely observed complex networks.PLoS computational biology · 2022Article
- Next generation single-molecule techniques: Imaging, labeling, and manipulation in vitro and in cellulo.Molecular cell · 2022Review
- Recent biomedical advances enabled by HaloTag technology.Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al · 2022Article
- Inherited nuclear pore substructures template post-mitotic pore assembly.Developmental cell · 2021Article
- Transient non-specific DNA binding dominates the target search of bacterial DNA-binding proteins.Molecular cell · 2021Article
- Review
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Authors and funding
9 authors at 2 institutions in 2 countries.
Funding
Abstract
Cell biology is increasingly dependent on quantitative methods resulting in the need for microscopic labelling technologies that are highly sensitive and specific. Whilst the use of fluorescent proteins has led to major advances, they also suffer from their relatively low brightness and photo-stability, making the detection of very low abundance proteins using fluorescent protein-based methods challenging. Here, we characterize the use of the self-labelling protein tag called HaloTag, in conjunction with an organic fluorescent dye, to label and accurately count endogenous proteins present in very low numbers (<7) in individual Escherichia coli cells. This procedure can be used to detect single molecules in fixed cells with conventional epifluorescence illumination and a standard microscope. We show that the detection efficiency of proteins labelled with the HaloTag is ≥80%, which is on par or better than previous techniques. Therefore, this method offers a simple and attractive alternative to current procedures to detect low abundance molecules.
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