Overview

  • Product name
    Acetylcholinesterase Assay Kit (Fluorometric -Red)
    See all Acetylcholinesterase kits
  • Detection method
    Colorimetric/Fluorometric
  • Sample type
    Serum, Plasma, Cell Lysate
  • Assay type
    Enzyme activity
  • Sensitivity
    0.1 mU/ml
  • Assay time
    0h 20m
  • Species reactivity
    Reacts with: Other species, Mammals
  • Product overview

    Acetylcholinesterase Assay Kit (Fluorometric -Red) ( ab138873) uses a red indicator to quantify the choline produced from the hydrolysis of acetylcholine by AChE through choline oxidase-mediated enzyme coupling reactions.


    The assay can be used for monitoring and quantifying AChE activity in blood, cell extracts or other solutions. The fluorescence intensity of the indicator is proportional to the amount of choline formed, which is proportional to the AChE activity.


    The assay is an optimized “mix and read” assay using a simple one-step fluorometric protocol to detect as little as 0.01 mU AChE in a 100 µl assay volume (0.1 mU/ml). The assay can be performed in a 96-well or 384-well microtiter-plate format, and can be easily read with a fluorescence microplate reader at Ex/Em = 540/590 nm or with an absorbance microplate reader at OD = 575 nm.


    Acetylcholinesterase assay protocol summary:
    - add samples and standards to wells
    - add assay mixture and incubate for 10-30 min
    - analyze with microplate reader

  • Notes

    Please note this product does not differentiate between acetylcholesterase (AchE) or butyrylcholinesterase (BChE) activity as both enzymes can hydrolyze acetylcholine.

    If you are looking for a colorimetric kit, we recommend Acetylcholinesterase Assay Kit (Colorimetric) (ab138871). If your plate reader has a filter to measure green fluorescnce (Ex/Em = 490/520 nm), you can use Acetylcholinesterase Assay Kit (Fluorometric - Green) (ab138872).

  • Platform
    Microplate reader

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 200 tests
    AbRed Indicator 1 vial
    Acetylcholine 1 vial
    Acetylcholinesterase Probe 2 units
    Acetylcholinesterase Standard 1 vial
    Assay Buffer 1 x 10ml
    Dilution Buffer 1 x 10ml
    DMSO 1 x 100µl
  • Research areas
  • Function
    Terminates signal transduction at the neuromuscular junction by rapid hydrolysis of the acetylcholine released into the synaptic cleft. Role in neuronal apoptosis.
  • Tissue specificity
    Isoform H is highly expressed in erythrocytes.
  • Sequence similarities
    Belongs to the type-B carboxylesterase/lipase family.
  • Cellular localization
    Cell membrane; Cell junction > synapse. Secreted. Cell membrane and Nucleus. Only observed in apoptotic nuclei.
  • Information by UniProt
  • Alternative names
    • ACEE
    • ACES_HUMAN
    • Acetylcholinesterase
    • AChE
    • Apoptosis related acetylcholinesterase
    • ARACHE
    • N ACHE
    • N-ACHE
    • YT
    • YT blood group
    see all

Images

  • AChE measured in cell lysates showing activity per 1 million cells

     

  • AChE measured in various species showing activity per mL sample tested

     

  • Acetylcholinesterase dose response was measured in a solid black 96-well plate with ab138873 using a fluorescence microplate reader. As low as 0.01 mU/well (0.1 mU/ml) acetylcholinesterase can be detected with 20 minutes incubation (n=3).

Protocols

References

ab138873 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abreviews
AChE Bestimmung im Zellkulturlysat
Acetylcholinesterase Assay Kit (Fluorometric Red) abcam #ab138873
Projekt: B7 Protokoll: Z1 Datum: 04.09.2015
Proben: Schafszellen K2 P7 (Bohrmehl)

Vorbereitung:
Allgemein:
• Ansetzen von AbRed Stock Solution: Zugabe von 40µL DMSO (Comp. G) in ein Tube mit AbRed Indikator (Comp.A), leicht vortexen,(Aliquotieren, bei -20°C lichtgeschützt einfrieren) =250x
• Ansetzen von AChE Standard: Zugabe von 100 µL Assay Puffer (Comp. E) in ein Tube mit AChE Standard (Comp.D), leicht vortexen, (Aliquotieren, bei -20°C einfrieren) =50 units/mL
• Ansetzen von ACh Stock Solution: Zugabe von 100 µL Assay Puffer (Comp. E) in ein Tube mit ACh (Comp. C), leicht vortexen, (Aliquotieren, bei -20°C einfrieren) =1000x Stock solution
• Ansetzen von AChE assay mixture: Zugabe von 5 mL Assay Puffer (Comp. E in ein Tube mit AChE Probe (Comp. B), vortexen, Aliquotieren, bei -20 °C einfrieren
• Zellen ( ab 10 000) in 250 µL Assay Puffer (Comp. E) resuspendieren, mit Sonifier homogenisieren (15 Impulse), anschließend mit 14 000 rpm 5 Minuten bei 4°C zentrifugieren


AChE Bestimmung:

• Ansetzen von AChE Standardlösungen:
1000 mU/mL AChE Standard Solution: 5 µL von 50 U/mL AChE Standard in ein Tube mit 245 µL Dilution Puffer (Comp. F) pipettieren, vortexen
100 mU/mL AChE Standardlösung: 25 µL 1000 mU/mL AChE Standard Solution zu 225 µL Dilution Puffer pipettieren, vortexen
30 mU/mL AChE Standardlösung:7,5 µL 1000 mU/mL AChE Standard Solution zu 243 µL Dilution Puffer pipettieren, vortexen
10 mU/mL AChE Standardlösung: 25 µL 100 mU/mL AChE Standardlösung zu
225 µL Dilution Puffer pipettieren, vortexen
3 mU/mL AChE Standardlösung: 25 µL 30 mU/mL AChE Standardlösung zu
225 µL Dilution Puffer pipettieren, vortexen
1 mU/mL AChE Standardlösung: 25 µL 10 mU/mL AChE Standardlösung zu
225 µL Dilution Puffer pipettieren, vortexen
0,3 mU/mL AChE Standardlösung: 25 µL 3 mU/mL AChE Standardlösung zu
225 µL Dilution Puffer pipettieren, vortexen
0,1 mU/mL AChE Standardlösung: 25 µL 1 mU/mL AChE Standardlösung zu
225 µL Dilution Puffer pipettieren, vortexen
• Ansetzen von ACh Probe mixture: Zugabe von 1,7 µL von 1000x ACh Stock Solution in ein Tube mit 1,7 mL AChE assay mixture, vortexen, Zugabe von 6,8 µL 250x AbRed Stock Solution, leicht vortexen (AChE Bestimmung)


Durchführung:

• 50 µL Zelllysat, Assay Puffer für Blank und AChE Standardlösung in 2x Bestimmung in die schwarze 96-Wellplatte pipettieren
• AChE spezifische Inhibitor: Donepezil hydrochlorid (SIGMA# D6821) 0,43 µL 23 mM Stock in 99,6 µL Wasser lösen, vortexen, 1 µL in den zu inhibierenden Well pipettieren (100 nM)
• Rivastigmin tartrat (SIGMA#SML0881) 0,32 µL 31,22 mM Stock in 99,7 µL Wasser lösen, vortexen, 1 µL in den zu inhibierenden Well pipettieren. (100 nM)
• 50 µL ACh Probe Mixture zu den Zelllysaten, AChE Standards, Zellysate mit Inhibitoren und Blank in 2x Bestimmung pipettieren
• Platte schütteln
• In Reader Fluoreszenz 530/590 Sensitivität 35 die Kinetik messen.
• Auswertung: 4- Parameter Kurve, x-Achse AChE Standard, y-Achse Blank Read nach1 h Inkubation.

Miss. Olga Dakischew

Verified customer

Submitted Oct 06 2015

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