Fluoro SSAO

$415.00

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Description

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 – Fluorescent Plate Reader.

Additional information

Kit Size

500

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 differs 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 activity is associated with blood vessels, mainly in smooth muscle layers, however, it is also associated with the 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 the membrane-bound form. The possible sources of plasma SSAO are still unclear, but it has been suggested that it may be derived from the 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 the 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.

Readable Documents

Title Name
Protocol Fluoro SSAO Protocol.pdf
Datasheet Fluoro SSAO Datasheet.pdf
msds msds Fluoro SSAO.pdf

Kit contents and storage

Part# Reagent Temperature
Part # 3019 5X Reaction Buffer pH 7.4, 1 Bottle 2-8°C
Part # 6005 Horseradish Peroxidase, 1 Vial 2-8°C
Part # 4008 Detection Reagent, 1 Vial -20°C (Aliquot in Single Use Vials)
Part # 7001 SSAO Substrate Benzylamine, 1 Vial -20°C (Aliquot in Single Use Vials)
Part # 6006 SSAO Enzyme, 1 Vial -20°C (Aliquot in Single Use Vials)
Part # 7003 Pargyline: Monoamine Oxidase B Inhibitor(9-11), 1 Vial -20°C (Aliquot in Single Use Vials)
Part # 7004 Semicarbazide-Sensitive Amine Oxidase Inhibitor, 1 Vial -20°C (Aliquot in Single Use Vials)

References

Yayin Tahiri Kasim et.al., Turk J Biochem; 29 (3);247-254 (2004); Semicarbazide-Sensitive Amine Oxidase: Biochemical and Physiological Properties
B.A.Callingham,A.Holt,J.Elliott.J.Neural Transm.(Suppl.)32 ,279 –290 (1990).
B.A.Callingham and M.A.Barrand.J.Neural Transm.(Suppl.)23 ,37 –54 (1987).
G.A.Lyles.J.Neural Transm.(Suppl.)41 ,387 –396 (1994).
K.Magyar, Z.Mészáros and P.Mátyus . Semicarbazide-sensitive amine oxidase.Its physiological significance Pure Appl.Chem.,Vol.73,No.9,pp.1393 –1400,2001.
Boomsma F.,van Dijk J.,Bhaggoe U.M.,Bouhuizen A.M.B.,van den Meiracker A.H.(2000)Variation in semicarbazide-sensitive amine oxidase activity in plasma and tissues of mammals.Comp.Biochem.
Ekblom J.,Gronval J.L.,Garpenstrand H.,Nillson S.,Oreland L.(2000)Is semicarbazide-sensitive amine oxidase in blood plasma partly derived from the skeleton?.Neurobiology,8,129-135.
McGuirl M.A., Dooley D.M.(1999)Copper-containing oxidases. Curr.Opin.Chem.Biol.,3,138-144.
Methods Enzymology 142, 617 (1997).
Holt A. ; Sharman D.F. ; Baker G.B. ; Palcic M.M. A Continuous Spectrophotometric Assay for Monoamine Oxidase and Related Enzymes in Tissue Homogenates Analytical Biochemistry, January 1997, vol. 244, no. 2, pp. 384-392(9).
Biochem Pharmacol 18, 1447 (1969).

 

Reaction

Benzylamine + O2+ H2O + SSAO arrow Benzaldehyde + NH3 + H202

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

Excitation:530-571nm

Emission: 590-600nm

flurescent_semicarbazide

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.