3,3',5,5' - Tetramethylbenzidine as an Ames Test Negative Chromogen for Horse-Radish Peroxidase in Enzyme-Immunoassay

J Immunoassay. 1981;2(3-4):187-204. doi: 10.1080/15321818108056977.


The use of 3,3',5,5' - tetramethylbenzidine as non-mutagenic chromogen for the end point determination in enzyme-immunoassay (EIA) is described. In sandwich EIAs for HCG and HBsAg and in a competitive EIA for testosterone, the colour yield with TMB was superior to that obtained with o-phenylene diamine (OPD), which was by far the best chromogen for horse-radish peroxidase until now. This led to an improvement of sensitivity and precision of the assays and makes EIA even more competitive with other types of immunoassays.

MeSH terms

  • Animals
  • Benzidines / pharmacology*
  • Cattle
  • Chorionic Gonadotropin / analysis
  • Chromogenic Compounds / pharmacology*
  • Female
  • Hepatitis B Surface Antigens / analysis
  • Horseradish Peroxidase / metabolism*
  • Immunoenzyme Techniques
  • Mutagenicity Tests
  • Peroxidases / metabolism*
  • Rabbits
  • Solubility
  • Testosterone / analysis


  • Benzidines
  • Chorionic Gonadotropin
  • Chromogenic Compounds
  • Hepatitis B Surface Antigens
  • 3,3',5,5'-tetramethylbenzidine
  • Testosterone
  • Horseradish Peroxidase
  • Peroxidases