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Localization of the two major allergens in rye-grass pollen using specific monoclonal antibodies and quantitative analysis of immunogold labelling

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Summary

The intracellular localization of the two major allergens, Lol p I and Lol p IX, in rye-grass anthers was examined using monoclonal antibodies FMCA1 (specific for Lol p I) and FMCA7 (specific for Lol p IX) with immunocytochemical techniques and quantitative analysis. A newly developed anhydrous fixation technique in a mixture of glutaraldehyde, paraformaldehyde and 2, 2-dimethoxypropane followed by embedding in LR Gold resin resulted in both improved infiltration of pollen grains compared with existing techniques and the localization of these water-soluble antigens in their original sites compared with diffusion artefacts following aqueous methods. After anhydrous fixation, Lol p I was predominantly located in the electron-opaque regions of the cytosol of the vegetative cell of the tricellular pollen grains (24 counts μm-2), whereas Lol p IX was detected mainly within starch granules (16 counts μm-2). For both Lol p I and Lol p IX, similar labelling was detected in the cells of the endothecium and middle layer (18 counts μm-2), but none was found in the tapetal cells or orbicules.

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Authors and Affiliations

  1. School of Botany, University of Melbourne, 3052, Parkville, Victoria, Australia

    P. E. Taylor, M. B. Singh & R. B. Knox

  2. Botany Department, LaTrobe University, 3083, Bundoora, Victoria, Australia

    I. A. Staff

Authors
  1. P. E. Taylor
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  2. I. A. Staff
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  3. M. B. Singh
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  4. R. B. Knox
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Taylor, P.E., Staff, I.A., Singh, M.B. et al. Localization of the two major allergens in rye-grass pollen using specific monoclonal antibodies and quantitative analysis of immunogold labelling. Histochem J 26, 392–401 (1994). https://doi.org/10.1007/BF00160051

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  • DOI: https://doi.org/10.1007/BF00160051

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