Monoamine Oxidase B (MAO-B) Human Enzymatic LeadHunter Assay - TW

Procedure Summary:

Human recombinant MAO-B expressed in insect cells is used. Test compound and/or vehicle is preincubated with 2.29 µg/ml enzyme‡ in phosphate buffer pH 7.4 for 15 minutes at 37°C. The reaction is initiated by addition of 50 µM kynuramine for 60 minute incubation period and terminated by further addition of 1.2 N NaOH. Determination of the amount of 4-hydroxyquinoline formed is read spectrofluorimetrically. Compounds are screened at 10 µM.

Substrate:

50 µM Kynuramine

Incubation:

60 min at 37°C

Control Inhibitor:

R(-)-Deprenyl

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2