Commercial polyclonal and monoclonal histostaining PAP kits. Immunoperoxidase reagents and performance characteristics in comparison with self-prepared immunoreagents

Abstract

Universal, polyclonal and monoclonal immunoperoxidase staining kits from BioGenex, Dako and Ortho were employed for the localization of antigens such as gastrin, prostate specific antigen, IgA, IgG, AFP and CEA in histological sections from formaldehyde fixed and paraffin embedded human specimens. The kit components were controlled by immunohistological and serological assays and were also compared with self-prepared reagents. In connection with specific primary antibodies, universal/basic kits gave reliable localization of defined antigens. The optimal concentration of the primary antibodies had to be established by dilution experiments. In the case of polyclonal kits, typical antigen localization was obtained in selected tissue sections with all the respective kits. CEA kits also stained strongly NCA molecules present in organs such as colon, stomach and liver. BioGenex polyclonal kits gave almost stronger stainings than kits from Dako and Ortho. Irrespective of which kit from different commercial sources is used, development of peroxidase activity with AEC/H2O2 often had to be stopped far below the recommended incubation time of 40 min or overstaining with color change from reddish to muddy green occurred. The latter was attributed to insufficiently balanced kit reagents, an interpretation which was supported by quantitative serological studies. Sensitivity of immunohistological reactivity was much enhanced by pretreatment of tissue sections with Pronase. Thus, stronger immunostainings and larger numbers of positive cells were detected than in conventionally rehydrated sections.(ABSTRACT TRUNCATED AT 250 WORDS)