[The proteolytic pretreatment of formalin-fixed tissue in immunohistochemical diagnosis].
Proteolytic enzymes were tested for improving histochemical localization of tissue antigens. Sections, 2-4 micron in thickness, were prepared on sodium-silicate coated slides from formalin-fixed, paraffin-embedded human biopsies. A modification of the Sternberger technique (PAP) and the indirect immunofluorescence method were used for the localization of 15 various antigens: heavy chain immunoglobulins, light chain immunoglobulins, alpha 1-fetoprotein, alpha 1-antichymotrypsin, myoglobin, fibronectin, factor VIII (ass. ag), fibrinogen, lysozyme and cytokeratin. The ability of different proteolytic enzymes (trypsin, pronase, pepsin) to unmask antigen in formalin-fixed sections were tested by variation of concentration, incubation time, temperature and pH. Although proteolytic unmasking to some extent is reliable, good restoration of antigenicity is not always possible. Best results were obtained with pronase E (Serva, FRG).
Bahn
H
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Institut für Pathologische Anatomie des Bereiches Medizin der Martin-Luther-Universität Halle-Wittenberg.
D000818:Animals; D000941:Antigens / Q000032:analysis*; D005353:Fibronectins / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007128:Immunoglobulin Fragments / Q000032:analysis; D007633:Keratins / Q000032:analysis; D009113:Muramidase / Q000032:analysis; D009211:Myoglobin / Q000032:analysis; D010434:Pepsin A / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D011402:Pronase / Q000378:metabolism; D011817:Rabbits; D013696:Temperature; D014357:Trypsin / Q000378:metabolism; D000514:alpha 1-Antichymotrypsin / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis
D000941:Antigens; D005353:Fibronectins; D007128:Immunoglobulin Fragments; D009211:Myoglobin; D000514:alpha 1-Antichymotrypsin; D000509:alpha-Fetoproteins; D005557:Formaldehyde; D007633:Keratins; D009113:Muramidase; D010447:Peptide Hydrolases; D014357:Trypsin; D010434:Pepsin A; D011402:Pronase
Zur proteolytischen Vorbehandlung formalinfixierter Gewebe in der immunohistochemischen Diagnostik.