The Histochemical Journal

, Volume 13, Issue 3, pp 461–480 | Cite as

Immunofluorescent localization of two water-soluble glycoproteins including the major allergen from the pollen of ryegrass,Lolium perenne

  • B. J. Howlett
  • H. I. M. V. Vithanage
  • R. B. Knox


Two major glycoproteins have been localized in sectioned grains of ryegrass pollen by direct and indirect immunofluorescence methods using Fluorescein isothiocyanate (FITC)-labelled IgC fractions of antisera. These glycoproteins are the major allergen Group 1 allergen, and a principal antigen Antigen A. Four methods of fixation were employed: freeze-drying, methanol, 2.5% glutaraldehyde and 4% paraformaldehyde for 1 h at 4°C. The post-embedding staining technique of immunocytochemistry was used: anthers were embedded directly, or after dehydration, in JB-4 plastic resin and antibody reacted with sectioned pollen.

The effects of these fixatives on the antibody combining sites of the antigens were quantified by a solid phase radioimmunoassay using [125I]protein A to measure antibody binding. Glutaraldehyde was the only fixative to significantly depress antibody binding of both Antigen A and Group 1 allergen to their homologous antisera. This radioimmunoassay was modified to reyeal that FITC conjugation to either antibody did not impair antigen binding. In mature pollen, these antigens were located in the cytoplasm and in the complex wall. In developing grains early in the maturation period, specific fluorescence was concentrated at the periphery of the cytoplasm.


Fluorescein Isothiocyanate Antibody Binding Mature Pollen Antigen Binding Maturation Period 
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Copyright information

© Chapman and Hall Ltd 1981

Authors and Affiliations

  • B. J. Howlett
    • 1
  • H. I. M. V. Vithanage
    • 1
  • R. B. Knox
    • 1
  1. 1.School of BotanyUniversity of MelbourneParkvilleAustralia

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