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Cat. No.: HRP-100 (for 100mg)

Cat. No.: HRP-1k (for 1g)

Cat. No.: HRP-10k (for 10g)

Description

Horseradish Peroxidase (HRP), also known as Peroxidase (POD) from Horseradish, is an oxidoreductase enzyme containing heme that is extracted from the horseradish plant. It has the EC number 1.11.1.7 and the CAS number 9003-99-0. HRP catalyzes the oxidation of various organic and inorganic substrates using hydrogen peroxide or other organic peroxides as electron acceptors. The enzyme activity of this product is ≥300 U/mg.

HRP has a wide range of applications. Due to its small and stable molecular weight, as well as low non-specific binding due to glycosylation modifications, HRP is commonly used for labeling secondary antibodies, primary antibodies, Streptavidin, and other molecules for Western blotting, ELISA, immunohistochemistry, and other detection methods. HRP is also frequently used in biochemical assays.

This product is a high-purity HRP powder with a brown color. The brown color is due to the presence of heme cofactors in HRP and not due to impurities. The product has an RZ value ≥3, enzyme activity ≥340 U/mg, and solubility ≥10 mg/ml. RZ (Reinheitszahl) is defined as the ratio of the maximum absorbance at 403 nm (corresponding to heme content) to the maximum absorbance at 275 nm (corresponding to deheme HRP). The RZ value only reflects the heme content and does not represent the enzyme activity. The RZ value for high-purity HRP should be around 3.0. The enzyme activity unit is defined as the amount of enzyme required to oxidize 1.0 mg of pyrogallol to purpurogallin in 20 seconds at 20°C and pH 6.0.

HRP is a single-chain polypeptide containing four disulfide bonds. It is a glycoprotein formed by the combination of a colorless enzyme protein and a brown ferroprotoporphyrin. It has a molecular weight of approximately 44 kDa, including a peptide chain (33890 Da), heme (approximately 700 Da), and carbohydrate substances (approximately 9400 Da). The carbohydrate content is about 18%. Different HRP isoenzymes contain different types of carbohydrates, including galactose, arabinose, xylose, trehalose, mannose, glucosamine, and galactosamine. At least seven HRP isoenzymes have been discovered, with isoelectric points ranging from approximately 3.0 to 9.0.

The mechanism of action of HRP involves the binding of HRP with hydrogen peroxide to form the [HRP-H2O2] complex, which catalyzes the oxidation of various hydrogen donor substrates. The hydrogen donor substrates are often colorless reduced chromogenic substrates (such as DAB, TMB, ABTS), chemiluminescent substrates (such as luminol), and fluorogenic substrates (such as Amplex Red). The reaction generates colored dyes, chemiluminescence, or fluorescent products, which can be qualitatively or quantitatively detected using appropriate instruments.

HRP is commonly conjugated to antibodies using methods such as glutaraldehyde crosslinking mediated by disulfide bonds, periodate oxidation, or direct coupling through amino and sulfhydryl groups. Compared to β-galactosidase and alkaline phosphatase, HRP has a smaller molecular weight and higher stability, making it the most ideal antibody label among the three.

HRP is most active at a pH range of 6.0-6.5 and remains stable within a pH range of 5.0-9.0. Common inhibitors of HRP include sodium azide, cyanides, L-cysteine, chromate salts, hydroxylamine compounds, vanadate esters, p-aminobenzoic acid, as well as metal ions such as Cd2+, Co2+, Cu2+, Fe2+, Mn2+, Ni2+, and Pb2+.

Precautions

• Please use this product immediately upon receipt or store it according to the recommended conditions in the instruction manual. With prolonged storage time, there may be a certain degree of decline in enzyme activity.
• This product is intended for scientific research use by professionals only. It is not intended for clinical diagnosis or treatment, and should not be used in food or pharmaceuticals. It should not be stored in a regular residential setting.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling this product.

Storage

Store at 4°C or -20°C in a light-protected manner. The product is valid for at least two years under these storage conditions.

Only for research and not intended for treatment of humans or animals

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