Overview

  • Product name
    Acetylcholinesterase Assay Kit (Colorimetric)
    See all Acetylcholinesterase kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture extracts, Cell Lysate, Tissue Lysate
  • Assay type
    Enzyme activity
  • Sensitivity
    1 mU/ml
  • Assay time
    0h 30m
  • Product overview

    Acetylcholinesterase Assay Kit ab138871 provides a convenient colorimetric assay for the detection of AChE activity in blood, cell extracts and other samples from mammals and other species.


    The acetylcholinesterase assay protocol uses DTNB to quantify the thiocholine produced from the hydrolysis of acetylthiocholine by AChE. The absorption intensity of DTNB adduct (410 nm) is used to measure the amount of thiocholine formed, which is proportional to the AChE activity.


    The assay will detect as little as 0.1 mU AChE in a 100 µL assay volume (1 mU/mL).  


    Acetylcholinesterase assay protocol summary:
    - add standards and samples to wells
    - add acetylthiocholine reaction mix and incubate for 10-30 min at room temp
    - analyze with a microplate reader

  • Notes

    Please note this product does not differentiate between acetylcholesterase (AchE) or butyrylcholinesterase (BChE) activity as both enzymes can hydrolyze acetylcholine. However, an acetylcholinesterase inhibitor like Donepezil hydrochloride can be used as a control.

    If you are looking for a fluorometric kit, we recommend Acetylcholinesterase Assay Kit (Fluorometric-Green) (ab133872) (Ex/Em = 490/520 nm) or Acetylcholinesterase Assay Kit (Fluorometric-Red) (ab133873) (Ex/Em = 540/590 nm).

  • Platform
    Microplate reader

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 200 tests
    Acetylcholinesterase Standard 1 x 5 units
    Acetylthiocholine 1 vial
    Assay Buffer 1 x 25ml
    DTNB 1 vial
  • 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

  • ab138871 was used to determine AChE activity in the cortex and in the hippocampus. AChE activity was measured at 410 nm.

  • AChE measured in mouse tissue lysates showing quantity (mU) per mg protein of tested sample

  • AChE measured in cell lysates showing quantity (mU) per 1 mln of tested cells

  • AChE measured in biological fluids showing quantity (mU) per mL of tested sample

  • Acetylcholinesterase dose response was measured in a white/clear bottom 96-well plate with ab138871 Acetylcholinesterase Assay Kit (Colorimetric) using a microplate reader. As low as 0.1 mU/well of acetylcholinesterase can be detected with 30 minutes incubation (n=3)

Protocols

References

This product has been referenced in:
  • Mendivil-Perez M  et al. iPSCs-derived nerve-like cells from familial Alzheimer's disease PSEN 1 E280A reveal increased amyloid-beta levels and loss of the Y chromosome. Neurosci Lett 703:111-118 (2019). Read more (PubMed: 30904577) »
  • Darwiche W  et al. Impact of chronic exposure to the pesticide chlorpyrifos on respiratory parameters and sleep apnea in juvenile and adult rats. PLoS One 13:e0191237 (2018). Rat . Read more (PubMed: 29357379) »
See all 10 Publications for this product

Customer reviews and Q&As

1-5 of 5 Q&A

Answer

Our Mammalian Cell Lysis Buffer 5X (ab179835) would be the best lysis buffer to use.

<1%Chaps also is OK. We have not checked NP40 and RIPA, but they should be OK theoretically.

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Question
Answer

Our recommended buffer for preparation of cell or tissue lysates for the Acetylcholinesterase assay ab138871 is ab179835.

https://www.abcam.com/index.html?datasheet=179835

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Question
Answer

We didn't test the inhibitory effect of ethanol on acetylcholinesterase, but the publications below showed that ethanol will inhibit activity of AChE.

http://www.ncbi.nlm.nih.gov/pubmed/3767274
http://link.springer.com/article/10.1023%2FA%3A1007555924661

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Answer



The recommended protocol to prepare whole blood for measuring Acetylcholinesterase is as follows:

Whole blood and red blood cells (RBC) lysates can be prepared by diluting 1.0 ml of blood or RBC in 19.0 ml of 0.03% Triton X-100 in water. And then diluted in dilutions in Assay Buffer for further experiment.

All of the information we have for each of our products is listed on the online datasheet for your convenience. These are updated as soon as any new information is brought to our attention. Therefore, unfortunately, there aren’t any references from other customers, at least that we know, using this kit with rat blood.

However, please bear in mind that the Abpromise® guarantee ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 12 months of purchase.

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Question
Answer

Gracias por contactarnos.
Me confirman mis compañeros del laboratorio que el kit es válido para realizar unos 200 ensayos, teniendo en cuenta un volumen aproximado de muestra por ensayo de 50uL.
Espero que esta información sea útil. En caso contrario, no dudes en contactarnos de nuevo.

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