Horseradish peroxidase

high-resolution terminology - matching measurements at high-resolution

Horseradish peroxidase

Description

Horseradish peroxidase readily combines with hydrogen peroxide (H₂O₂) and the resultant [HRP-H₂O₂] complex can oxidize a wide variety of hydrogen donors.

Abbreviation: HRP

MitoPedia methods: Fluorometry 

Application in HRFR

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):

1. Weigh 3,33 mg
2. Dissolve in 1 mL MiR05.
3. 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.

Measurement of H₂O₂ with Amplex UltraRed

Reaction

Donor + H₂O₂ → Oxidized Donor + 2 H₂O

Known inhibitors

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.