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Fluoro SSAO

Fluoro SSAO Cell Technology Fluorescent Enzymatic Assays
Fluoro SSAO
Fluorescent Semicarbazide-Sensitive Amine Oxidase Detection Kit

$395.00

in stock
Product code: SSAO100
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Product 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.

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 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.

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).

Kit contents and Long Term storage

ReagentPart#Temperature
5X Reaction Buffer pH 7.4, 1 BottlePart # 30192-8C
Horseradish Peroxidase, 1 VialPart # 60052-8C
Detection Reagent, 1 VialPart # 4008-20C (Aliquot in Single Use Vials)
SSAO Substrate Benzylamine, 1 VialPart # 7001-20C (Aliquot in Single Use Vials)
SSAO Enzyme, 1 VialPart # 6006-20C (Aliquot in Single Use Vials)
Pargyline: Monoamine Oxidase B Inhibitor(9-11), 1 VialPart # 7003-20C (Aliquot in Single Use Vials)
Semicarbazide-Sensitive Amine Oxidase Inhibitor, 1 VialPart # 7004-20C (Aliquot in Single Use Vials)
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