 |
| 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
|
|
|
| Fluorescent
Enzymatic Assays |
|
|
| Mitochondria
Membrane Potential detection |
|
|
| aCella -
Bioluminescence Assays Kits |
|
|
| Cytotoxicity |
|
|
| ELISA
Products |
|
|
| Cathepsin B,K,L
Detection Kits |
|
|
|
|
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| |
|
Fluoro
AChE™
Fluorescent
Acteylcholinesterase
(AChE) Detection Kit - Detection of AChE in RBC, Saliva, and Tissue
Lysates
|
| Key
Benefits: |
- Non
Radioactive assay can monitor multiple time points to follow kinetics.
- One-step,
no wash assay
- Versatile:
Can Detect AChE activity in RBC's, Saliva and Tissue Lysates
- Readout
- 96 well Fluorescence Plate Reader
|
| Introduction
|
Exposure to
chemical nerve agents,
pesticides and certain drugs (anesthetics, cocaine and therapeutical
drugs) reduces the activity of red blood cell (RBC)
acetylcholinesterase (AChE). The RBC-AChE can be used as a biomarker to
monitor suppressed and or increased AChE function in the peripheral and
central nervous system (9).
Acetylcholinesterase
(AChE) is one of the
most important enzymes involved in nerve transmission. The enzyme is
bound to cellular membranes of excitable tissue (synaptic junction,
endoplasmic reticulum,
etc) 1-3.
Acute toxicity to humans and animals through inhibition of AChE by
both nerve gases and an important class of pesticides
has long been a field of intensive
scientific investigation 4,5.
AChE inhibitors have also been used clinically as Alzheimer’s
treatments (e.g.,
tacrine (tetrahydroaminoacridine)) 6
and are the subject of
increasing interest
in various disease processes and treatment strategies 7,8.
However, both environmental detection of AChE inhibitors and
development of modulators of AChE enzymatic activity as drugs have been
hampered by the difficulty and complexity of the current assay methods.
|
| |
| Assay
Principle |
| We have developed a highly sensitive, very
rapid, extremely simple assay to determine acetylcholinesterase
activity in RBC’s, using the natural substrate,
acetylcholine. Additionally, by using specific inhibitors, the kit can
be used to detect AChE activity in a variety of samples. A series of
coupled enzyme reactions quickly translates the presence of active AChE
into a change in the fluorescence of a quenched detection reagent.
AChE + ATP + H20
+
coupled enzyme reaction + quenched dye ----> Fluorescent
Dye (Ex:530-570nm
Em:590-600nm
|
| Citations |
Gonadal hormones modulate the potency of the disruptive effects of
donepezil in male rats responding under a nonspatial operant learning
and performance task -
Leonard, Stuart T.; Hearn, John K.; Catling, Andrew D.; Winsauer, Peter
J - Behavioral Pharmacology Feb 2010
|
| References |
-
Politoff,
A., Blitz, A., and Rose, S.:
Incorporation of Acetylcholinesterase Into Synaptic Vesicles is
Associated with Blockade of Synaptic Transmission, Nature
256, 324, 1975
-
Friedenberg,
R., and Seligman, A.:
Acetylcholinesterase at the Myoneural Junction: Cytochemical
Ultrastructure and Some Biochemical Considerations, J Histochem Cytochem
20, 771, 1972
-
Nachmansohn,
D.: Proteins in Excitable
Membranes, Science 168,
1059, 1970.
-
HA Berman and MM Decker.
Kinetic, equilibrium,
and spectroscopic studies on dealkylation ("aging") of alkyl
organophosphonyl acetylcholinesterase.
Electrostatic control of enzyme topography.
J. Biol. Chem., Aug 1986; 261: 10646-10652 .
-
Arie Ordentlich et al. The Architecture of
Human Acetylcholinesterase
Active Center Probed by Interactions with Selected Organophosphate
Inhibitors.
J. Biol. Chem., May 1996; 271: 11953-11962.
-
Levy R. Tetrahydroaminoacridine
and Alzheimer's disease. Lancet, 1987 Feb 7;1(8528):322.
-
Bolognesi ML et al. Propidium-based
polyamine ligands as potent inhibitors of acetylcholinesterase and
acetylcholinesterase-induced amyloid-beta aggregation. J Med Chem. 2005
Jan 13;48(1):24-7.
-
Schallreuter KU et al. Activation/deactivation
of acetylcholinesterase by H2O2: more evidence for oxidative stress in
vitiligo. Biochem Biophys Res Commun. 2004 Mar 5;315(2):502-8.
- Nigg HN,
Knaak JB. Blood cholinesterases as human
biomarkers of
organophosphorus pesticide exposure. Rev. Environ. Contam. Toxicol,
2000:163: p29-111.
|
| Kit
contents (for 100 assays) |
- Component
A: Detection Reagent Diluent,
5.5mL.........Part# 3023
- Component
B: Coupled enzyme reagent,
5.5mL..........…Part# 3024
- ComponentC:
1vial detection
reagent………………………………Part#
4016
- Component
D: 5X Reaction
Buffer………………………………………Part#
3011
- Component
E: Acetylcholine…………………………………………………Part#
7012
- Component
E: Red Blood Cell
Acetylcholinesterase………Part# 6020
|
| The
following kits are
available: |
Catalog
#
|
Size
|
Price
(US$) |
| AChE
100-2 |
100
Tests |
$395 |
| AChE
100-3 |
500
Tests |
$1595 |
|
| * Please call
888 7 ASSAYS (888-727-7297)
or email info@celltechnology.com
for volume pricing |
|
| |
 Product Data Sheet |
| Protocol |
| |
|
Time=10
min
|
Dilution
|
Volume
used
|
ug
protein/well
|
RFU *
|
|
RBC
|
1:1000
|
10uL
|
5
|
3450
|
|
RBC no
Ach
|
1:1000
|
10uL
|
5
|
152
|
|
Rat
brain
|
1:500
|
10uL
|
20
|
5864
|
|
Rat
brain no Ach
|
1:500
|
10uL
|
20
|
296
|
|
Saliva
|
neat
|
10uL
|
ND
|
925
|
|
Saliva
no Ach
|
neat
|
10uL
|
ND
|
772
|
|
|
|
|
