Peroxidase Inactivated

Description

• Application: Inactivated peroxidase is used to study regeneration of inactivated peroxidase enzymes[1]. It has been used to study the mechanism of peroxidase solubilization[2] and it is used to study antitumor activity of immobilized peroxidase systems[3]. It may be used for developing differential staining techniques in nerve tissue preparations.
• Biochem/physiol Actions: HRP readily combines with hydrogen peroxide (H2O2) and the resultant [HRP-H2O2] complex can oxidize a wide variety of hydrogen donors. The optimal pH is 6.0-6.5 and the enzyme is most stable in the pH range of 5.0-9.0. HRP can be conjugated to antibodies by several different methods that include the use of glutaraldehyde, periodate oxidation, disulfide bonds, and also via amino- and thiol-directed cross-linkers. It is smaller and more stable than the enzyme labels, β-galactosidase and alkaline phosphatase. Hence, it is the most desired label. Also, its glycosylation leads to lower non-specific binding.[4] It is also used for the determination of glucose[5] and peroxides[6] in solution. Sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and ions such as Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, and Pb2+ are found to inhibit the enzyme activity.[7]
• Unit Definition: One unit will form 1.0 mg purpurogallin from pyrogallol in 20 sec at pH 6.0 at 20°C.
• Preparation Note: Prepared from Type VI horseradish peroxidase by a modification of the method of Teale.