Advantages of detecting monoclonal antibody binding to tissue sections with biotin and avidin reagents in Coplin jars.
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 biotinylated 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.
J M
JM
Bindl
R A
RA
Warnke
D000818:Animals; D000911:Antibodies, Monoclonal; D001360:Avidin; D001710:Biotin; D006041:Goats; D006735:Horseradish Peroxidase / Q000378:metabolism; D006736:Horses; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000032:analysis; D008722:Methods; D051379:Mice; D010047:Ovalbumin / Q000031:analogs & derivatives; D014066:Palatine Tonsil / Q000473:pathology; D013194:Staining and Labeling / Q000295:instrumentation*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D001360:Avidin; D001710:Biotin; D010047:Ovalbumin; D006735:Horseradish Peroxidase