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redcoon België GENTAUR BVBA

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1040 Brussels BELGIUM

  Tel + 32 16 58 90 45 

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75005 Paris, France

 Tel 01 43 25 01 50

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52074 Aachen, Germany

Tel  0241 56 00 99 68                   

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KOMABIOTECH
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Sacace Biotechnologies S.r.l.
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SCETI K.K
BIOSCIENCE
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EY Laboratories, Inc. Headquarters
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EXBIO Praha, a.s.
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252 42  Vestec
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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 

Kit contents (for 100 assays)
  1. Component A: Detection Reagent Diluent, 5.5mL.........Part# 3023
  2. Component B: Coupled enzyme reagent, 5.5mL..........…Part# 3024
  3. ComponentC: 1vial detection reagent………………………………Part# 4016
  4. Component D: 5X Reaction Buffer………………………………………Part# 3011
  5. Component E: Acetylcholine…………………………………………………Part# 7012
  6. 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

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