Table 10.5 Advantages of the Amplex UltraRed reagent over chromogenic reagents.

Peroxidase Detection Reagent Abs * Em * Notes
Amplex UltraRed reagent568581
  • More resistant to oxidation
  • More sensitive
  • Fast fluorescence (and color) development
Amplex Red reagent570585
  • Resistant to oxidation
  • Sensitive
  • Fast fluorescence (and color) development
ABTS (2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid))405NA
  • Readily oxidized
  • Slow color development
OPD (o-phenylenediamine)492NA
  • Light sensitive
TMB (3,3',5,5'-tetramethylbenzidine)450 (stopped reaction)
653 (kinetic assay)
  • Readily oxidizes
  • Precipitates easily due to low water solubility
* Absorption (Abs) and fluorescence emission (Em) maxima, in nm. NA = Not applicable.