Monoamine oxidase A&B detection

Spherotech, Inc.
27845 Irma Lee Circle, Unit 101
Lake Forest, IL 60045

Exalpha Biologicals, Inc.
2 Shaker Road, Unit B101
Shirley, MA 01464

SCETI K.K
BIOSCIENCE Export DF Kasumigaseki Place,3-6-7 Kasumigaseki Chiyoda-ku, Tokyo 100-0013 JAPAN

GENTAUR BVBA

VAT BE0473327336

Av. de l Armee 68 B4

1040 Brussels BELGIUM

Tel + 32 16 58 90 45

Fax + 32 16 50 90 45

GENTAUR France SARL

SIRET 48423788800017

Rue Lagrange, 9

75005 Paris, France

Tel 01 43 25 01 50

Fax 01 43 25 01 60

GENTAUR Germany Marienbongard 20

52074 Aachen, Germany

Tel 0241 56 00 99 68 Fax 0241 56 00 47 88

GENTAUR Pol Sp. Z.o.o. Ulica Ogarna 15/19B m2

80-826 GDANSK

Tel 00 48 58 760 77 08

Fax: 00 32 16 50 90 45

GENTAUR Italy

23015 Milano, Italy

Tel 02 36 00 65 93

Fax 02 36 00 65 94

Česká republika Praha
+420246019719

Danmark

+4569918806

Finland Helsset
+358942419041

Ελλάς Αθήνα
+302111768494

Ireland Dublin
+35316526556

Luxembourg
+35220880274

Magyarország Budapest
+3619980547

Nederland
+31208080893

Norge Oslo
+4721031366

Österreich
+43720880899

Sverige Stockholm
+46852503438

Schweiz Züri
+41435006251

Northern America

Canada Montreal
+15149077481

US New York
+17185132983

Monoamine oxidase (MAO) is a flavin-containing enzyme that catalyses the oxidation of a variety of amine-containing neurotransmitters such as serotonin, norepinephrine, epinephrine and dopamine to yield the corresponding aldehydes (1). MAO exists in two isoforms, namely MAO-A and MAO-B, which are the products of two distinct genes (2).

MAO-A and B exhibit different specificities to substrates and inhibitor selectivities. Extensive studies have been preformed to characterize their properties (3-7). MAO-A acts preferentially on serotonin and norepinephrine, and is inhibited by clorgyline. MAO-B acts preferentially on 2-phenylethylamine and benzylamine and is inhibited by deprenyl and pargyline.

Localized in the outer mitochondrial membrane, these enzymes are found throughout the body. Often only one form of the enzyme is present in a specific organ and/or within a specific cell type (8-9). In addition to their role in regulating neurotransmitters, these enzymes are also involved in processing biogenic amines (10) including tyramine (11).

The Fluoro MAO-A/B detection kit utilizes a non - fluorescent substrate, 10-Acetyl-3, 7-dihydroxyphenoxazine (ADHP) to detect H₂0₂ released from the conversion of a substrate to its aldehyde via MAO-A/B. Furthermore, H₂0₂ oxidizes ADHP in a 1:1 stoichiometry to produce a fluorescent product resorufin. This oxidation is catalyzed by Peroxidase.

H₂0₂ + Detection reagent (non-fluorescent) + Peroxidase ---------> Resorufin (fluorescent)

Figure1. Recombinant MAO-B was serially diluted in 1X Reaction Buffer. 100 mL of diluted MAO-B per well was mixed with 100mL of Reaction Cocktail. The reaction was incubated at Room Temperature in the dark for 1 hour. Fluorescence was measured with a microplate reader using excitation at 530mM and fluorescence emission at 590mM.