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Recipe for fluorescent antibodies

Immunofluorescence

Nature Cell Biology volume 11, pages S10–S11 (2009)Cite this article

The idea to make antibodies visible was conceived to visualize group A haemolytic streptococci in the Aschoff nodules that characterize rheumatic fever. Initially, Coons and his colleagues repeated the work of John Marrack, who had successfully modified antibodies with tetrazotized benzidine to produce coloured antibodies without disturbing their antigen-binding properties. These coloured antibodies were too faint to be detected in dilute solutions, so Coons et al. turned to a fluorescent compound, anthracence isothiocyanate. They obtained brightly fluorescent antibodies that could agglutinate bacteria, indicating that the conjugation had not affected the antibodies' binding properties. However, when they tested the fluorescence in tissue sections, they discovered that human and mouse tissues autofluoresce at the same wavelength as anthracence.

...the basic idea remains unchanged...

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References

ORIGINAL RESEARCH PAPERS

  1. Coons, A. H., Creech, H. J. & Jones, R. N. Immunological properties of an antibody containing a fluorescent group. Proc. Soc. Expt. Biol. Med. 47, 200–202 (1941)

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  2. Coons, A. H., Creech, H. J., Jones, R. N. & Berliner, E. The demonstration of pneumococcal antigen in tissues by the use of fluorescent antibody. J. Immunol. 45, 159–170 (1942)

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FURTHER READING

  1. Coons, A. H. & Kaplan, M. H. Localization of antigen in tissue cells: improvements in a method for the detection of antigen by means of fluorescent antibody. J. Exp. Med. 91, 1–13 (1950)

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  2. Coons, A. H. The beginnings of immunofluorescence. J. Immunol. 87, 499–503 (1961)

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  1. Chief Editor, Nature Reviews Microbiology,

    Christiaan van Ooij

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  1. Christiaan van Ooij
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van Ooij, C. Recipe for fluorescent antibodies. Nat Cell Biol 11 (Suppl 1), S10–S11 (2009). https://doi.org/10.1038/ncb1932

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