Native Peroxidase from Horseradish

Products

Native Peroxidase from Horseradish

Product Specifications

Cat#	NATE-1015
Descriptions	High-activity Horseradish-derived Peroxidase catalyzes hydrogen peroxide-mediated chromogenic or chemiluminescent signal amplification across all mainstream diagnostic platforms. It serves as a universal labeling and signal development enzyme for clinical immunoassays, ELISA, chemiluminescence detection and colorimetric biochemical kits, offering high signal sensitivity and consistent batch activity for diagnostic reagent manufacturers.
Applications	This product can be served as core catalytic reagent for enzymatic quantitative detection of hydrogen peroxide in clinical diagnosis.
Synonyms	Donor: hydrogen-peroxide oxidoreductase
EC Number	EC 1.11.1.7
Catalysis	Donor + H₂O₂→ Oxidized donor + 2H₂O
Activity	≥ 180 U/mg (Peroxidase activity)
Appearance	Light brown to reddish brown powder, lyophilized
Additive	Not added
Molecular Weight	41,000 (SDS-PAGE)
Isoelectric Point	6.2, 7.2, 8.8
Optimum pH	6.0
pH Stability	6.0-10.0 (30°C, 30 min)
Optimum Temperature	40°C
Thermal Stability	up to 60°C (pH 6.0, 1 h)
Inhibitors	NaN₃, KCN
Unit Definition	1 unit is defined as the enzyme quantity which decomposes 1 µ mole of hydrogen peroxide per minute under the conditions described below.
Storage	24 months from the date of analysis when stored at 5°C or below in a dry place under sealed conditions.

General Properties

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Assay Principle

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Activity Assay

Unit Definition

1 unit is defined as the enzyme quantity which decomposes 1 µ mole of hydrogen peroxide per minute under the conditions described below.

Reagents

A. 4-Aminoantipyrine solution (4 mg/mL deionized water)
B. Triton X-100 solution (50 mg/mL deionized water)
C. Phenol solution (50 mg/mL deionized water)
D. 0.1 M Phosphate buffer (KH2PO4-NaOH, pH 7.0)
E. Phenol-buffer solution: Weigh 1.36g of KH2PO4 and dissolve in 80 mL of deionized water. Add 3 mL of Phenol solution (C) and 3 mL of Triton X-100 solution (B), then adjust the pH to 7.0 with 4N NaOH. Fill up to 100 mL with deionized water. This reagent should be used after incubation at 25℃ for 24 hours. (Can be used for 14 days at 25℃)
F. Substrate solution: Dilute 0.175 mL of hydrogen peroxide (35.5 % concentration) with deionized water and fill up to 200 mL.
G. Enzyme solution: Weigh out Peroxidase and dissolve in chilled 0.1 M Phosphate Buffer (D). Enzyme solution should be prepared so that the value of ΔOD/minute becomes in the range of 0.030±0.005.

Procedure

Pipette 2.0 mL of Phenol-buffer solution (E), 1 mL of Substrate solution (F), and 0.1 mL of 4-Aminoantipyrine solution (A) respectively into quartz cell (d=10 mm). Keep at 37±0.5℃ for 10 min. Then, pipette 0.1 mL of Enzyme solution (G) into the quartz cell and mix well immediately. Keep the reaction mixture at 37±0.5℃. Exactly at 2 minutes and 5 minutes after the addition of Enzyme solution (G), measure the absorbances of the reaction mixture at 500 nm (A2 and A5). As a blank, pipette 0.1M Phosphate buffer (D) into another quartz cell instead of the Enzyme solution (G) and follow the same procedure described above (Ab2 and Ab5).

Calculation

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Safe Handling Avoid inhalation; in the event of direct skin or eye contact, immediately flush the affected area with abundant water, and consult the SDS for further information.