Skip to main content
SpringerLink
Log in

Immunogold labelling is a quantitative method as demonstrated by studies on aminopeptidase N in microvillar membrane vesicles

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

Microvillar membrane vesicle preparations with varying content of aminopeptidase N were prepared from enterocytes of the pig small intestine. Postembedding immunogold labelling of aminopeptidase N was performed on these glutaraldehyde/paraformaldehyde-fixed, osmium tetroxide-treated and Epon-embedded microvillar membrane vesicles. The number of gold particles per micrometre microvillar membrane (labelling intensity) was calculated and compared to the corresponding enzymatic activity. A very close relationship was found between labelling intensity and aminopeptidase N activity, demonstrating that postembedding immunogold labelling can be used quantitatively.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Faulk, W. P. &Taylor, G. M. (1971) An immunocolloid method for the electron microscope,Immunochemistry 8, 1081–3.

    PubMed  Google Scholar 

  • Griffiths, G. &Hoppeler, H. (1986). Quantitation in immunocytochemistry: correlation of immunogold labeling to absolute number of membrane antigens.J. Histochem. Cytochem. 34, 1389–98.

    PubMed  Google Scholar 

  • Hansen, G. H., Sjöström, H., Norén, O. &Dabelsteen, E. (1987) Immunomicroscopic localization of aminopeptidase N in the pig enterocyte. Implications for the route of intracellular transport.Eur. J. Cell Biol. 43, 253–9.

    PubMed  Google Scholar 

  • Hansen, G. H., Norén, O. &Sjöström, H. (1988). Biosynthesis of microvillar proteins: Subcellular localization of intestinal aminopeptidase N along the crypt-villus axis. InMammalian Brush Border Membrane Proteins (edited byLentze, M. J. &Sterchi, E. E.) pp. 29–42. New York, Stuttgart: George Thieme Verlag.

    Google Scholar 

  • Hansen, G. H., Danielsen, E. M., Sjöström, H. &Norén, O. (1989) Organelles involved in the intracellular transport of newly synthesized aminopeptidase N and their acidity.Eur. J. Cell Biol. 49, 154–61.

    PubMed  Google Scholar 

  • Howell, K. E., Reuter-Carlson, U., Devaney, E., Paul Luzio, J. &Fuller, S. D. (1987) One antigen, one gold? A quantitative analysis of immunogold labeling of plasma membrane 5′-nucleotidase in frozen thin sections.Eur. J. Cell Biol. 44, 318–27.

    PubMed  Google Scholar 

  • Kessler, M., Acuto, O., Storelli, C., Murer, H., Müller, M. &Semenza, G. (1978). A modified procedure for the rapid preparation of efficiently transporting vesicles from small intestinal brush border membranes. Their use in investigating some properties of D-glucose and choline transport systems.Biochim. Biophys. Acta 506, 136–54.

    PubMed  Google Scholar 

  • Ottersen, O. P. (1989). Postembedding immunogold labelling of fixed glutamate: an electron microscopic analysis of the relationship between gold particle density and antigen concentration.J. Chem. Neuroanat. 2, 57–66.

    PubMed  Google Scholar 

  • Posthuma, G., Slot, J. W. &Geuze, H. J. (1987) Usefulness of the immunogold technique in quantitation of a soluble protein in ultra-thin sections.J. Histochem. Cytochem. 35, 405–10.

    PubMed  Google Scholar 

  • Reynolds, E. A. (1963). The use of lead citrate at high pH as an electron-opaque stain in electron microscopy.J. Cell Biol. 17, 208–12.

    PubMed  Google Scholar 

  • Sjöström, H., Norén, O., Jeppesen, L., Staun, M., Svensson, B. &Christiansen, L. (1978) Purification of different amphiphilic forms of a microvillus aminopeptidase from pig small intestine using immunoadsorbent chromatography.Eur. J. Biochem. 88, 503–11.

    PubMed  Google Scholar 

  • Sjöström, H. &Norén, O. (1982). Changes of the quaternary structure of microvillus aminopeptidase in the membrane.Eur. J. Biochem. 122, 245–50.

    Google Scholar 

  • Slot, J. W. &Geuze, H. J. (1985). A new method of preparing gold probes for multiple-labeling cytochemistry.Eur. J. Cell Biol. 38, 87–93.

    PubMed  Google Scholar 

  • Wang, B.-L. &Larsson, L.-I. (1985). Simultaneous demonstration of multiple antigens by indirect immunofluorescence or immunogold staining: novel light and electron microscopical double and triple staining method employing primary antibodies from the same species.Histochemistry 83, 47–56.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry C, The Panum Institute, University of Copenhagen, Blegdamsvej 3c, 2200, Copenhagen N, Denmark

    Gert H. Hansen, Lise-Lotte Wetterberg, Hans Sjöström & Ove Norén

Authors
  1. Gert H. Hansen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lise-Lotte Wetterberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hans Sjöström
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ove Norén
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hansen, G.H., Wetterberg, LL., Sjöström, H. et al. Immunogold labelling is a quantitative method as demonstrated by studies on aminopeptidase N in microvillar membrane vesicles. Histochem J 24, 132–136 (1992). https://doi.org/10.1007/BF01047462

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01047462

Keywords

Search

Navigation