 |
| Caspase / Apoptosis
Detection |
 |
High Throughput Screening
- Apo3HTS |
|
Caspase Poly,1,2,3,7,8,9,10
Detection - FAM-FMK |
|
Caspase Poly, 1,2,3,8,9,10,
Detection
- (SR-FMK) |
|
Antibody Specific Caspase 3 Detection
- Apo Active 3 (FITC) |
|
Antibody Specific Caspase 3 Detection
- Apo Active 3 (PE) |
|
| NOS/ROS Detection Kits |
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| Fluorescent Enzymatic
Assays |
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| Mitochondria
Membrane Potential detection |
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| aCella - Bioluminescence
Assays Kits |
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| Cytotoxicity |
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| ELISA
Products |
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| Cathepsin B,K,L
Detection Kits |
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|
Fluoro MAO™
Fluorescent
Monoamine Oxidase A&B Detection Kit
|
| Key Benefits: |
- Non Radioactive
- Can monitor multiple time points to
follow kinetics.
- One-step, no wash assay.
- Adaptable for High Throughput format
- Sensitive
- Applications - Standard
Fluorescence Plate Reader
|
| Assay Principle |
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.
|
| Reaction: |
| Substrate +
O2 + H202 + MAO-A/B ----------> aldehyde + NH3
+ H202 |
H202 +
Detection reagent (non-fluorescent) + Peroxidase --------->
Resorufin (fluorescent)
|
| The fluorescent monoamine
oxidase detection kit can be used to monitor MAO activity and screen of
MAO inhibitors. |
| References |
-
Waldmeier PC (1987)
Amine oxidases and their endogenous substrates. J Neural Transm Suppl
23:55–72.
-
Bach, A. W. J., N. C.
Lan, D. L. Johnson, C. W. Abell, M. E. Bembenck, S. W. Kwan, P. H.
Seeburg & J. C. Shih: cDNA clon-ing of human liver monoamine
oxidase A and B: molecular basis of differences in enzymatic
properties. Proc. Nat. Acad. Sci. U.S.A. 1988, 85, 4934–4938.
-
Johnston, J. P.: Some
observations upon a new inhibitor of mono-amine oxidase in brain
tissue. Biochem. Pharmacol. 1968, 17, 1285–1297.
- Suzuki, O., E. Noguchi & K. Yagi: A
simple fluorometric assay for type B monoamine oxidase activity in rat
tissues. J. Biochem. 1976, 79, 1297–1299.
-
Fowler, C. J. & B.
A. Callingham: Substrate-selective activation of rat liver
mitochondrial monoamine oxidase by oxygen. Biochem. Pharmacol. 1978,
27, 1995–2000.
-
Tipton, K. F.:
Enzymology of monoamine oxidase. Cell Biochem. Funct. 1986, 4,
79–87.
-
Youdim, M. B. H. &
M. Tenne: Assay and purification of liver monoamine oxidase. Methods
Enzymol. 1987, 142, 617–626.
-
Trendelenburg U,
Cassis L, Grohmann M and Langeloh A (1987) The functional coupling of
neuronal and extraneuronal transport with intracellular monoamine
oxidase. J Neural Transm Suppl 23:91–101.
- Yu PH, Davis BA and Boulton AA (1992)
Neuronal and astroglial monoamine oxidase: Pharmacological implications
of specific MAO-B inhibitors. Prog Brain Res 94:309–315.
- Strolin Benedetti M and Tipton KF (1998)
Monoamine oxidases and related amine
-
oxidases as phase I
enzymes in the metabolism of xenobiotics. J Neural Transm Suppl
52:149–171.
-
Hauptmann N, Grimsby
J, Shih JC and Cadenas E (1996) The metabolism of
tyramine by monoamine
oxidase A/B causes oxidative damage to mitochondrial DNA. Arch Biochem
Biophys 335:295–304.
Methods Enzymology:
Metabolism of Aromatic Amino Acids and Amines. Volume 142, page 617
(1997).13. 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).
-
D. W. R. Hall, Bridget
W. Logan and G. H. Parsons. Further studies on the inhibition of
monoamine oxidase by M & B 9302 (clorgyline)—I .Substrate
specificity in various mammalian species. Biochemical Pharma
|
| Kit
contents (for 500 assays) |
Description
|
Part#
|
Storage
after opening Kit or Reconstitution of Reagents. |
Bottle: 5X Reaction
Buffer pH 7.4
|
3020 |
4-80C |
| 1 vial: Detection
reagent |
4009
|
Aliquot in single use
vials: Below –200C
|
| 1 vial: Horseradish
Peroxidase |
6007 |
4-80C |
1 vial: MAO-B substrate
Benzylamine
|
7006 |
Aliquot in vials: Below
–200C
|
| 1 vial: MAO-A/B
substrate: Tyramine |
7005 |
Aliquot in single use
vials: Below –200C
|
| 1 vial Pargyline:
Monoamine Oxidase B inhibitor (12-14) |
7003 |
Aliquot in vials: Below
–200C
|
| 1 vial Clorgyline:
Monoamine Oxidase A inhibitor(12-14) |
7002 |
Aliquot in single use
vials: Below –200C
|
| The
following kits are available: |
Catalog #
|
Size
|
Price (US$) |
FLMAO 100-3
|
500 Tests |
$245 |
|
| * Please call 888 7 ASSAYS
(888-727-7297) or email info@celltechnology.com
for volume pricing |
|
|
