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| Caspase
/ Apoptosis
Detection |
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High
Throughput Screening
- Apo3HTS |
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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) |
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Antibody
Specific Caspase 3 Detection
- Apo Active 3 (PE) |
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Detection Kits |
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| Fluorescent
Enzymatic Assays |
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| Mitochondria
Membrane Potential detection |
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| Cytotoxicity |
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| ELISA
Products |
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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 |
$225 |
|
| * Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for
volume pricing |
|
|
