Back
Spectrophotometric quantification of horseradish peroxidase with o-phenylenediamine.
Journal article

Spectrophotometric quantification of horseradish peroxidase with o-phenylenediamine.

  • Fornera S Department of Materials, ETH Zürich, CH-8093 Zürich, Switzerland.
  • Walde P
  • 2010-08-10
Published in:
  • Analytical biochemistry. - 2010
Horseradish Peroxidase
Hydrogen Peroxide
Hydrogen-Ion Concentration
Phenazines
Phenylenediamines
Spectrophotometry
English The assay conditions for the spectrophotometric quantification of horseradish peroxidase (HRP) with the chromogenic substrate o-phenylenediamine (OPD) at 25°C in the presence of hydrogen peroxide (H(2)O(2)) were optimized. With [OPD](0)=3.14 mM and [H(2)O(2)](0)=80 μM at pH 7.2, the initial formation of only one of the possible reaction products and intermediates, 2,3-diaminophenazine (DAP, λ(max)=417 nm), was observed. The rate of DAP formation during the first 30 min of reaction was followed spectrophotometrically and found to linearly depend on the HRP concentration between 5 and 45 pM under the conditions used.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/183607
Statistics
Document views: 21 File downloads: