Semicarbazide sensitive amine oxidase detection

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Fluoro SSAO
Fluorescent Semicarbazide-Sensitive Amine Oxidase Detection Kit

Key Benefits:

Non Radioactive
Can monitor multiple time points to follow kinetics.
Monitors Enzymatic Activity
One-step, no wash assay.
Adaptable for High Throughput format
Enzyme Positive Control included in Kit
Applications - Fluorescence Plate Reader

Assay Principle

Semicarbazide-sensitive amine oxidase (SSAO) is a common name for a widely distributed enzyme in nature. In man this enzyme is present in the vascular system and circulates in plasma. SSAO differ from the monoamine oxidases A and B in substrate and inhibitor patterns. These enzymes have been widely studied and their tissue distribution, molecular properties, substrate specificities and inhibitor sensitivities are extensively reviewed (2,3).

SSAO exists in two forms: tissue bound and soluble (plasma SSAO). Tissue bound SSAO acitvity is associated with blood vessels, mainly in smooth muscle layers, however it is also associated with spleen, placenta, bone marrow, kidney, sclera, retina, endothelial cells, adipocytes, chondrocytes and fibroblasts. (4,5). Evidence suggests that Plasma SSAO originates from the cleavage of membrane-bound form. The possible sources of plasma SSAO are still unclear, but it has been suggested that it may be derived from liver, retina, placenta and bone tissue (6,7,8).

SSAO’s functional role has been suggested to be involved in: apoptosis, atherogenesis, cell adhesion, leucocyte trafficking, glucose transport and local production of hydrogen peroxide. Reports of elevated levels of SSAO have been reported in congestive heart failure, diabetes mellitus, alzheimer’s disease and various other inflammatory diseases (1).

Furthermore byproducts of SSAO deamination, such as formaldehyde and methylglyoxal, have been proposed to be involved in pathogenesis of cancer, aging and atherosclerosis.

The Fluoro SSAO detection kit utilizes a non- fluorescent detection reagent, to detect H₂0₂ released from the conversion of Benzylamine to Benzaldehyde via SSAO. Furthermore H₂0₂ oxidizes this detection reagent in a 1:1 stoichiometry to produce a fluorescent product resorufin. This oxidation is catalyzed by Peroxidase.

Reaction:

Benzylamine + O2+ H2O + SSAO

Benzaldehyde + NH3 + H₂0₂

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

Excitation:530-571nm
Emission: 590-600nm

Kit contents (for 500 assays)
Description	Part	Storage after opening Kit or Reconstitution of Reagents.
1 Bottle: 5X Reaction Buffer pH 7.4	3019	4-8⁰C
1 vial: Detection reagent for 500 Assays	4008	Aliquot in single use vials: Below –20⁰C
1 vial: Horseradish Peroxidase	6005	4-8⁰C
1 vial: SSAO substrate Benzylamine	7001	Aliquot in vials: Below –20⁰C
1 vial: SSAO Enzyme	6006	Aliquot in single use vials: Below –20⁰C
1 vial Pargyline: Monoamine Oxidase B inhibitor ^(9-11)	7003	Aliquot in vials: Below –20⁰C
1 vial Semicarbazide: Semicarbazide-sensitive amine oxidase inhibitor.	7004	Aliquot in single use vials: Below –20⁰C

The following kits are available:
Catalog	Size	Price (Eur €)
SSAO 100-3	500 Tests	€345

Figure 1. Bovine serum Semicarbazide-sensitive amine oxidase was serially diluted in 1X Reaction buffer. The serially diluted samples were run as described in the protocol. The samples were read after a 3 hours incubation period. Excitation: 530nm and emission: 590nm.