Advantages of detecting monoclonal antibody binding to tissue sections with biotin and avidin reagents in Coplin jars

Am J Clin Pathol. 1986 Apr;85(4):490-3. doi: 10.1093/ajcp/85.4.490.

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 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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Avidin*
  • Biotin*
  • Goats
  • Horseradish Peroxidase / metabolism
  • Horses
  • Humans
  • Immunoglobulin Fab Fragments / analysis
  • Methods
  • Mice
  • Ovalbumin* / analogs & derivatives
  • Palatine Tonsil / pathology
  • Staining and Labeling / instrumentation*

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Fab Fragments
  • Avidin
  • Biotin
  • Ovalbumin
  • Horseradish Peroxidase