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Immunogold probes for light and electron microscopic localization of Ole e I in several Oleaceae pollens

MC Fernandez, A Olmedilla, JD Alche, P Palomino, C Lahoz and MI Rodriguez-Garcia

Departamento de Bioquimica, Bologia Celular y Molecular de Plantas, Estacion Experimental del Zaidin, CSIC, Granada, Spain.

We investigated the immunolocalization of the olive major allergen Ole e I and Ole e I-like proteins in pollen from several Oleaceae species [olive (Olea europaea), ash (Fraxinus excelsior), privet (Ligustrum vulgaris), lilac (Syringa vulgare), and forsythia (Forsythia suspensa)]. Crossreactions among different pollens were found in enzyme immunoassays. For immunolocalization with light microscopy we used the silver enhancement technique with three monoclonal antibodies (1D8, 10H1, and 16G2) that recognize three different epitopes of the allergen Ole e I. Our findings show that the silver enhancement technique is very useful when several antibodies are to be used for rapid screening of different materials. MAb 10H1 gave the most precise results and was selected for further immunolocalization studies with transmission electron microscopy. The epitope recognized by this MAb was localized exclusively in the endoplasmic reticulum in olive pollen. In lilac, privet, and ash pollen, most of the reactivity was also seen in the endoplasmic reticulum; however, the 10H1 epitope was not detected in forsythia pollen.

Volume 44, Issue 2, pp. 151-158, 02/01/1996
Copyright © 1996 by The Histochemical Society


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 J. Cell Sci.Home page
J de Dios Alche, A. Castro, A Olmedilla, M. Fernandez, R Rodriguez, M Villalba, and M. Rodriguez-Garcia
The major olive pollen allergen (Ole e I) shows both gametophytic and sporophytic expression during anther development, and its synthesis and storage takes place in the RER
J. Cell Sci., January 8, 1999; 112(15): 2501 - 2509.
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