Horseradish peroxidase: Difference between revisions

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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 200µl into 500µl 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 ^[1]: sodium azide, cyanide, L-cystine,dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd²⁺, Co²⁺, Cu²⁺, Fe³⁺, Mn²⁺, Ni²⁺, and Pb²⁺ ions.

References

↑ Zollner H (1993) Handbook of Enzyme Inhibitors, 2nd ed, Part A:367-8.

[1] Zollner H (1993) Handbook of Enzyme Inhibitors, 2nd ed, Part A:367-8.

[1]

@@ Line 12: / Line 12: @@
 ::: '''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 200µl into 500µl Eppendorf tubes.
 :::: The use of horseradish peroxidase type II in fluorometric measurement of peroxide production is recommended by [[Tretter L|Laszlo Tretter]].