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| Product Name | Peroxidase Activity Colorimetric Assay Kit (Guaiacol Substrate) |
| Catalog No. | ETR-HMM-0083 |
| Description | Colorimetric assay for peroxidase (POD) activity using guaiacol as the reducing substrate and hydrogen peroxide as the oxidant. Peroxidase catalyzes the oxidation of guaiacol to tetraguaiacol (brown-orange chromophore, λmax = 470 nm) in the presence of H₂O₂. The simple, rapid protocol is ideal for measuring peroxidase activity in plant tissues, horseradish peroxidase (HRP) preparations, and enzyme engineering applications. |
| Intended Use | Peroxidase activity measurement in plant tissue extracts; HRP conjugate activity verification; enzyme kinetics characterization; food science peroxidase inactivation studies; environmental monitoring of peroxidase-based bioremediation. |
| Principle / Technology | H₂O₂ + guaiacol (reduced, colorless) → 2 H₂O + tetraguaiacol (oxidized, brown-orange) catalyzed by peroxidase; color development at 470 nm is proportional to enzyme activity |
| Detection Method | Colorimetric; kinetic or endpoint at 470 nm; typical assay volume 200-1,000 µL |
| Sample Type | Plant tissue extracts, HRP preparations, cell culture supernatants; 10-100 µL sample per assay |
| Sensitivity / LOD | LOD 0.01 U/mL peroxidase; linear range 0.01-5 U/mL in kinetic mode; 1 U = formation of 1 mg tetraguaiacol per minute |
| Specificity | Specific for heme-containing peroxidases (HRP, plant peroxidases, lactoperoxidase); non-heme peroxidases (glutathione peroxidase) do not efficiently utilize guaiacol; catalase interference minimal at recommended H₂O₂ concentration |
| Reaction Conditions / Protocol | Mix sample, guaiacol substrate (0.2% v/v), H₂O₂ (0.015% v/v) in phosphate buffer pH 6.0; monitor A470 increase kinetically for 3-5 min; calculate activity from initial linear rate using ε470 = 26.6 mM⁻¹cm⁻¹ for tetraguaiacol |
| Components / Formulation | Guaiacol substrate (liquid, 1% v/v in ethanol), hydrogen peroxide solution (0.3% v/v, stabilized), assay buffer (sodium phosphate 50 mM, pH 6.0), HRP positive control (lyophilized, 10 U), tetraguaiacol standard (for endpoint calibration, optional) |
| Storage Conditions | 2-8 °C for all components; H₂O₂ protected from light; guaiacol solution in tightly sealed bottle (volatile) |
| Shelf Life | 12 months from manufacture date |
| Package Specifications | 200 assays, 500 assays, 1,000 assays |
| Product Form | Liquid reagents; lyophilized positive control |
| Key Features | Classic guaiacol method — literature-standard peroxidase assay; rapid kinetic protocol (3-5 min); absorbance-based — no specialized equipment; HRP positive control and tetraguaiacol standard for calibration; applicable to diverse sample types |
| Purity | Guaiacol ≥99% by GC; H₂O₂ concentration verified by permanganate titration; HRP positive control RZ >3.0 |
| Concentration | As specified per kit; calibrator and substrate concentrations traceable to reference |
| Activity / Unit Definition | One unit defined as the amount of enzyme that catalyzes conversion of 1 µmol substrate per minute under specified conditions |
| Molecular Weight | Varies by target enzyme as documented |
| Source / Origin | Recombinant or purified from natural sources; synthetic chromogenic/fluorogenic substrates |
| pH Range / Optimal pH | pH 6.0–9.0 depending on specific enzyme assay |
| Shipping Conditions | Ambient; guaiacol solution is flammable — ship per Class 3 regulations |
| Expiration Date / Stability | 12 months at 2-8 °C; guaiacol stock stable when tightly sealed — evaporative loss changes concentration; H₂O₂ solution stable at 4 °C for 12 months; discard H₂O₂ if bubbles form in storage (decomposition) |
| Regulatory / Compliance | For laboratory and research use only; RUO; guaiacol is irritant and flammable; H₂O₂ is oxidizer; ISO 9001 |
| Compatibility | Compatible with standard microplate readers (absorbance, fluorescence, luminescence modes) |
| Recommended Buffer System | Tris or phosphate buffer optimized per enzyme assay |
| Application Notes / Precautions | Include positive control and inhibitor control in each assay; read within linear range; protect light-sensitive substrates |
| Batch-to-Batch Consistency | Enzyme activity and standard curve linearity within ±15% of reference lot |
For research use only, not for clinical use.
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