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:
  • 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) »
  • Kucharewicz K  et al. Simultaneous induction and blockade of autophagy by a single agent. Cell Death Dis 9:353 (2018). Read more (PubMed: 29500364) »
See all 9 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or 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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Zebrafish Samples

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We have tested this product with zebrafish brain and embryo lysates. A zebrafish mutant line sb55 (Behra et al., 2002) was used for the validation experiments as a control. In this mutant line homozygous mutants sb55-/- has abolished acetycholinesterase activity compared to wildype controls and homozygous mutation is lethal for embryos after 4 dpf due to paralysis. The measurement taken from homozygous mutants at 4dpf is approximately zero and there is a drastic difference between mutants and wildtypes at 4dpf in terms of acetylcholinesterase activity. In the adult fish mutation is not lethal for heterozygous mutants(sb55+/-) and heterozygous mutants have lower levels of brain acetylcholinesterase (Ninkovic et al., 2006). Our data is also confirming the previous observations by other groups working with zebrafish model and indicate that this product is giving quite accurate measurements with zebrafish brain and embryo samples.

Extraction Protocol: Brain tissues and embryos were homogenized in ice cold 100 ul of 25 mM Tris HCl pH:8. Then centrifuged at 13 000 rpm for 20 mins at 4°C. Collected supernatants were used for the assay.


Behra, M., Cousin, X., Bertrand, C., Vonesch, J.L., Biellmann, D., Chatonnet, A., and Strähle, U. (2002) Acetylcholinesterase is required for neuronal and muscular development in the zebrafish embryo. Nature Neuroscience. 5(2):111-118.

Ninkovic, J., Folchert, A., Makhankov, Y. V., Neuhauss, S. C., Sillaber, I., Straehle, U., & Bally‐Cuif, L. (2006). Genetic identification of AChE as a positive modulator of addiction to the psychostimulant D‐amphetamine in zebrafish. Journal of neurobiology, 66(5), 463-475.

Miss. Elif Karoğlu

Verified customer

Submitted May 10 2019

Abreviews
The kit is very easy to use, precise and fast.
I wanted to determine Exosomes presence and amount in the supernatant of cells and I was very satisfied with the result obtained with the kit.

Abcam user community

Verified customer

Submitted Apr 04 2017

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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