Skip to main content
Log in

Ultrastructural localization of human kidney antigens using monoclonal antibodies

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

Summary

A highly sensitive method of ultrastructural-immunoperoxidase staining was developed for use with monoclonal antibodies which have been raised in this laboratory to a variety of antigens of the human kidney. Because of the human kidney. Because of the susceptibility of the antigens to fixation and processing, a four layer, pre-embedding method of staining was used. Results confirmed and clarified previously reported light microscopy results, indicating that an antigen recognized by the PHM5 antibody was found on the podocyte cell membrane within the glomerulus and was not present within the glomerular basement membrane. The antigen was also present on the extraglomerular endothelial cell membrane. The study also demonstrated the presence of an antigen specific to endothelial cells throughout the renal cortex, and gave further insight into the precise localization of glomerular basement membrane components including fibronectin. The method of staining is now being used together with detailed ultrastructural studies to identify the cells produced from isolated glomeruli in tissue culture.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Avrameas, S. &Ternynck, T. (1971) Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penetration.Immunochemistry 8, 1175–9.

    Google Scholar 

  • Becker, G., Hancock, W. W., Karaft, N. &Atkins, R. C. (1981) Monoclonal antibodies to human macrophage and leucocyte common antigens.Pathology 13, 669–80.

    Google Scholar 

  • Courtoy, P. J., Kanwar, Y. S., Hynes, R. O. &Farquhar, M. G. (1980) Fibronectin localisation in the rat glomerulus.J. Cell Biol. 87, 691–6.

    Google Scholar 

  • Courtoy, P. J., Picton, D. H. &Farquhar, M. G. (1983) Resolution and limitations of the immunoperoxidase procedure in the localisation of extracellular matrix antigens.J. Histochem. Cytochem. 31, 945–51.

    Google Scholar 

  • Courtoy, P. J., Timpl, R. &Farquhar, M. G. (1982) Comparative distribution of laminin Type IV collagen and fibronectin in the rat glomerulus.J. Histochem. Cytochem. 30, 874–86.

    Google Scholar 

  • Glasgow, E. E., Hancock, W. &Atkins, R. C. (1981) The technique of glomerular culture. InTechniques for Nephropathology (edited byAllen, D. &Dowling, J. P.) Boca Raton: CRC Press pp. 87–93.

    Google Scholar 

  • Graham, R. C. Jr. &Karnovsky, M. J. (1964) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of the mouse kidney. Ultrastructural cytochemistry by a new technique.J. Histochem. Cytochem. 14, 291–302.

    Google Scholar 

  • Hancock, W. &Atkins, R. C. (1981) Monoclonal antibodies to human renal antigens.Aust. N. Z. Med. 11, 594.

    Google Scholar 

  • Hancock, W. &Atkins, R. C. (1983) Monoclonal antibodies to human glomerular cells: A marker for glomerular epithelial cells.Nephron 33, 83–90.

    Google Scholar 

  • Hancock, W., Becker, G. J. &Atkins, R. C. (1982) A comparison of fixatives and immunohistological technics for use with monoclonal antibodies to cell surface antigens.Am. J. Clin. Path. 78(6), 825–31.

    Google Scholar 

  • Hancock, W., Kraft, N., Clarke, F. &Atkins, R. C. (1984) Production of monoclonal antibodies to fibronectin, Type IV collagen and other antigens of the human glomerulus.Pathology 16, 197–206.

    Google Scholar 

  • Jackson, A. E., Glasgow, E. F. &Atkins, R. C. (1989) Ultrastructural immunoperoxidase investigations of human glomerular cells in tissue culture.Nephron 52, 62–71.

    Google Scholar 

  • Kraehenbuhl, J. P. &Jamieson, J. D. (1971) Localisation in intracellular antigens by immunoelectron microscopy.Int. Rev. Exper. Pathol. 13, 1–53.

    Google Scholar 

  • McLean, I. W. &Nakane, P. K. (1974) Periodate-Lysineparaformaldehyde fixative. A new fixative for immunoelectron microscopy.J. Histochem. Cytochem. 22, 1077–83.

    Google Scholar 

  • Martinez-Hernandez, A., Marsh, C. A., Clark, C. C., Macarak, E. J. &Brownell, A. G. (1981) Fibronectin: Its relationship to basement membranes. II. Ultrastructural studies in rat kidney.Coll. Res. 5, 405–18.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jackson, A.E., Atkins, R.C. & Glasgow, E.F. Ultrastructural localization of human kidney antigens using monoclonal antibodies. Histochem J 23, 509–516 (1991). https://doi.org/10.1007/BF01041177

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01041177

Keywords

Navigation