SKU: AFG-HZM-131-5KU

AffiZYME® Ascorbate oxidase

Vendor AffiGEN
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AffiZYME® Ascorbate oxidase

Abbreviation: ASOD

Preparation and Specification:

  • Appearance: Light blue amorphous powder, lyophilized
  • Activity: Grade III 200U/mg-solid or more (containing approx. 70% of stabilizers)
  • Contaminants: Catalase ≤1.0×10⁻¹ %, Phosphatase ≤2.0×10⁻² %
  • Stabilizers: BSA, borax, basic amino acids.

Properties:

  • Stability: Stable at -20°C for at least one year
  • Molecular Weight: 132,000, 140,000
  • Isoelectric Point: Between 6.0 and 7.8, 8.2
  • Michaelis Constant: 2.5×10⁻⁴M (Ascorbate)
  • Structure: 8 copper atoms per mol of enzyme
  • Inhibitors: Cyanide, Na₂S, diethyldithiocarbamate (Na)
  • Optimum pH: 5.6
  • Optimum Temperature: Approx. 30°C
  • pH Stability: pH 7.0−10.0 (25°C, 17hr)
  • Thermal Stability: Below 40°C (pH 8.0, 30min)
  • Substrate Specificity: Oxidizes ascorbic acid and several ascorbic derivatives

Applications:

  • Useful for enzymatic determination of ascorbic acid and eliminating interference in clinical analysis.

Assay:

  • Principle: Ascorbate oxidase catalyzes the oxidation of ascorbic acid to dehydroascorbic acid.
  • Unit Definition: One unit causes the decrease of one micromole of ascorbic acid per minute.
  • Method: Spectrophotometry at 245nm.
  • Reagents: Ascorbic acid solution, Na₂HPO₄ solution, HCl solution, Enzyme diluent.
  • Procedure: Reaction mixture preparation, enzyme addition, stop reaction with HCl, measure optical density.
  • Calculation: Activity calculation based on optical density changes.