POST-EMBEDDING IMMUNOLABELING - SOME EFFECTS OF TISSUE-PREPARATION ON THE ANTIGENICITY OF PLANT-PROTEINS

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  • 1 January 1982
    • journal article
    • research article
    • Vol. 28  (2) , 251-256
Abstract
The sensitivity of detection of proteins by immunolabeling embedded and sectioned material depends upon retention of antigenicity during tissue processing. Losses in pea [Pisum sativum cv. Greenfeast] seed storage protein antigenicity were assessed using a solid phase radioimmunoassay. Storage proteins adsorbed to wells of microtiter plates were treated with the various tissue preparation steps and reacted with 125I-antibodies to the storage proteins. The glutaraldehyde or formaldehyde fixation steps caused .apprx. 60% and 30% loss in antigenicity, respectively. With each subsequent preparative step, losses accumulate and following treatment with Spurr''s epoxy resin reached .apprx. 85% and 90%, respectively. If Lowicryl K4M, a methacrylate-acrylate embedding medium was used, losses were retained at .apprx. 70% and 60%, respectively. This was verified on sections of embedded material using 2 labeling procedures, with protein A-gold and ferritin as the markers.