The Histochemical Journal

, Volume 16, Issue 7, pp 771–787 | Cite as

Influence of decalcifying agents on immunoreactivity of formalin-fixed, paraffin-embedded tissue

  • John B. Matthews
  • Gillian I. Mason


The influence of eight decalcifying agents on the immunoreactivity of formalin-fixed, paraffin-embedded tissue for immunoglobulins, lysozyme, factor VIII-related antigen and keratin was studied using the unlabelled antibody peroxidase-antiperoxidase (PAP) method. Limited studies were also performed on tissues fixed in acid-formalin mixtures. All tissues were stained using an indirect immunoperoxidase method with mouse monoclonal antibodies to IgM, kappa and lambda light chains. The results suggest that, with controlled trypsinization of sections, it was possible to obtain optimal immunostaining for all tested antigens, with adequate preservation of histological structure, after decalcification in neutral EDTA or 10% aqueous acetic or formic acid. Tissue treated with agents containing mineral acids exhibited variable immunoreactivity and impaired counterstaining.


Acetic EDTA Monoclonal Antibody Formic Acid Lysozyme 
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Copyright information

© Chapman and Hall Ltd. 1984

Authors and Affiliations

  • John B. Matthews
    • 1
  • Gillian I. Mason
    • 1
  1. 1.Immunology Laboratory, Department of Oral Pathology, The Dental SchoolUniversity of BirminghamBirminghamUK

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