Superheating antigen retrieval

Anthony S.Y. Leong, Eung Seok Lee, Hui Yin, Megan Kear, Zenobia Haffajee, Debbie Pepperall

    Research output: Contribution to journalArticlepeer-review

    27 Citations (Scopus)


    Heat-induced antigen retrieval in a variety of solutions has been shown to enhance the immunoreactivity of a wide range of antigens in routine formalin-fixed, paraffin-embedded tissues. Accurate time and temperature control is important for standardization and optimization of the procedure but is difficult to achieve. This study used a device to attain precise time and temperature control for antigen retrieval at 120°C under 1.9 bar pressure. It compares the efficacy of this method with antigen retrieval in a conventional pressure cooker, by microwave heating at 98°C, and ultrasound retrieval at 40 and 70 W for 40 and 100 seconds. Multitissue and multitumor blocks containing a spectrum of normal tissues and a variety of tumors, respectively, were used, and 42 routine diagnostic antibodies were applied with a standard peroxidase conjugated streptavidin technique. Sections in which antigen retrieval was not performed served as controls. The three heat-induced methods showed distinctly better immunostaining for all antigens compared with those obtained with ultrasound retrieval. The latter method did not produce consistent staining and intensity, and the extent of staining was only marginally better than sections not subjected to antigen retrieval. Superheating at 120°C produced the best overall results with the exception of antibodies to cytokeratin clones Cam 5.2, AE1/3, and 34BE12 in which superheating resulted in slightly inferior immunostaining compared with heating in a pressure cooker and at 98°C.

    Original languageEnglish
    Pages (from-to)263-268
    Number of pages6
    JournalApplied Immunohistochemistry and Molecular Morphology
    Issue number3
    Publication statusPublished - 2002 Sept


    • Antigen retrieval
    • Diagnostic antibodies
    • Immunohistochemistry
    • Microwaves
    • Pressure cooker
    • Superheating
    • Ultrasonication

    ASJC Scopus subject areas

    • Pathology and Forensic Medicine
    • Histology
    • Medical Laboratory Technology


    Dive into the research topics of 'Superheating antigen retrieval'. Together they form a unique fingerprint.

    Cite this