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Monoamine oxidase (EC 1.4.3.4) is an enzyme composed of different polypeptides. Monoamine oxidation catalyzes the oxidative deamination of various biological amines in brain and peripheral tissues by producing hydrogen peroxide. Monoamine oxidase plays an important role in maintaining the regulation of synaptic transmission, emotional behavior and other brain functions.

Monoamine oxidase exists in several isoforms, the most typical of which are monoamine oxidase (MAO) A and MAO B. MAO A is generally inhibited by low concentrations of clorgyline, whereas MAO B is generally inhibited by low concentrations of I-deprenyl hydrochloride.

Monoamine Oxidase Assay

1. Solution Preparation Substrate

• 1.0% v/v (0.10 M) benzylamine in 67 mM potassium phosphate buffer, pH 7.2.
• Enzyme solution: Dissolve in 67 mM potassium phosphate buffer (pH 7.2) to a concentration of 10 mg/mL. Store at 2–8°C for a maximum of 1 week.

2. Assay Procedure

• Adjust the spectrophotometer to 250 nm and 25°C.
• Pipette 2.8 mL of 67 mM potassium phosphate buffer (pH 7.2) and 0.1 mL of substrate solution into each cuvette.
• Incubate in the spectrophotometer for 3–4 minutes to allow temperature equilibration.
• Add 0.1 mL of enzyme solution and record the increase in A250 over 6–8 minutes. Calculate ΔA250/min using the linear portion of the curve. A lag phase of 2–3 minutes may occur.

Room temperature in continental US; may vary elsewhere.

Store at -20°C for 2 years.