Horseradish peroxidase readily combines with hydrogen peroxide (H2O2) and the resultant [HRP-H2O2] complex can oxidize a wide variety of hydrogen donors.
Application in HRR
- HRP: Horseradish peroxidase (peroxidase from horseradish Type II, essentially salt-free, lyophilized powder, 150-250 units/mg solid); Sigma P 8250, store at 4 °C.
- Preparation of 500 U/mL stock solution (dissolved in MiR05):
- Weigh 3,33 mg
- Dissolve in 1 mL MiR05.
- Aliquot 200 µL into 500 µL Eppendorf tubes.
- The use of horseradish peroxidase type II in fluorometric measurement of peroxide production is recommended by Laszlo Tretter.
- Donor + H2O2 → Oxidized Donor + 2 H2O
- According to Sigma-Adrich, citing : sodium azide, cyanide, L-cystine,dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, and Pb2+ ions.
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- ↑ Zollner H (1993) Handbook of Enzyme Inhibitors, 2nd ed, Part A:367-8.
MitoPedia methods: Fluorometry