Advantages of Detecting Monoclonal Antibody Binding to Tissue Sections with Biotin and Avid in Reagents in Coplin® Jars

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Abstract
We describe a method of biotin/avidin-peroxidase detection using second and third stage reagents in Coplin® jars. This method allows a large quantity of sections to be stained simultaneously with a minimal amount of technical time involved. A wide range of mouse monoclonal antibodies of varying specificities and isotypes were used to stain both frozen and paraffin-embedded sections of various normal and neoplastic tissues. Three different biotiriylated anti-mouse antibodies were tested, including F(ab′)2 antibody fragments of one, followed by horseradish peroxidase conjugated avidin. All monoclonal antibodies employed gave good staining, using incubation times of 30–50 minutes. The staining was done during a mean period of 25 to 27 days with an average staining load of 500 sections per Coplin jar.