The use of 3,3',5,5'-tetramethylbenzidine as a peroxidase substrate in microplate enzyme-linked immunosorbent assay

J Immunoassay. 1987;8(1):29-41. doi: 10.1080/01971528708063053.


The assay conditions for the use of 3,3',5,5' tetramethylbenzidine (TMB) in microplate enzyme-linked immunosorbent assay are described. TMB is a safe (non-mutagenic) chromogen that is more sensitive than OPD as a substrate for horseradish peroxidase. We describe the optimum storage and assay conditions for this chromogen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Protozoan / analysis
  • Benzidines*
  • Chromogenic Compounds
  • Enzyme-Linked Immunosorbent Assay*
  • Giardia / immunology
  • Horseradish Peroxidase
  • Substrate Specificity


  • Antigens, Protozoan
  • Benzidines
  • Chromogenic Compounds
  • 3,3',5,5'-tetramethylbenzidine
  • Horseradish Peroxidase