• Product name
    Choline/Acetylcholine Assay Kit
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Cell culture extracts, Other biological fluids, Tissue Extracts
  • Assay type
  • Sensitivity
    > 0.01 nmol/well
  • Range
    0.01 nmol/well - 5 nmol/well
  • Assay time
    0h 40m
  • Species reactivity
    Reacts with: Mammals, Other species
  • Product overview

    Choline/Acetylcholine Assay Kit (ab65345) provides a simple and sensitive means for quantifying Choline and Acetylcholine by either a colorimetric or fluorometric method. In the assay free choline is oxidized to betaine, via the intermediate betaine aldehyde. The reaction generates products which react with the Choline Probe to generate color (λ= 570 nm), and fluorescence (Ex/Em 535/587 nm). Acetylcholine can be converted to choline by adding acetylcholinesterase to the reaction. The kit can detect choline and acetylcholine (total choline – free choline) in various biological samples such as in blood, cells, culture media, fermentation media, etc. There is no need for pretreatment or purification of samples. The kit can detect ~10 pmol-5 nmol of choline or acetylcholine.

    Cells should be cultured in choline chloride free medium.

    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Choline and acetylcholine play important roles in many biological processes. Choline is an essential nutrient, commonly grouped with the B complex vitamins, that plays key roles in many biological processes. Choline is a precursor for the synthesis of acetylcholine (AchE), a critical neurotransmitter in both the periferial and central nervous systems.

    If you need to purchase additional Choline Probe for this product, please see ab139505.



  • Acetylcholine levels from mouse brain tissue was measured using Acetylcholine assay kit (ab65345). Normal mice were anesthetized with thiopental sodium at 20minutes after the end of  electroacupuncture (EA) treatment to determine levels of ACh (Figure A). Immunohistochemical staining  mAChR (muscarinic acetylcholine receptor) M3 in mouse cerebral cortex tissue 20 minutes after the end of EA treatment (Image B).

  • Choline measured fluorometrically in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample

  • Choline measured fluorometrically in cell lysates showing quantity (pmol) per 1 mln of tested cells

  • Choline measured fluorometrically in biological fluids showing quantity (nmol) per mL of tested sample



This product has been referenced in:
  • Garrido-Gil P  et al. Aging-related dysregulation in enteric dopamine and angiotensin system interactions: implications for gastrointestinal dysfunction in the elderly. Oncotarget 9:10834-10846 (2018). Functional Studies . Read more (PubMed: 29541380) »
  • van Loo B  et al. Structural and Mechanistic Analysis of the Choline Sulfatase from Sinorhizobium melliloti: A Class I Sulfatase Specific for an Alkyl Sulfate Ester. J Mol Biol 430:1004-1023 (2018). Read more (PubMed: 29458126) »

See all 10 Publications for this product

Customer reviews and Q&As

J’ai contacté le laboratoire concernant les limites de détection de ce kit et de sa sensibilité.
Comme montré sur la courbe standard pour la détection colorimétrique, la limite de détection inf...

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I suggest running a time course to see how much time is needed for your samples to show a linear increase and then plateau off. It is recommended to choose the final incubation time within this linear increase regime.
Also, please note it is impor...

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ab65345 cannot be used for in-cell assay. It is meant for cell lysates.

Thank you for your email and your interest in our products.

I can confirm that you are right. It will be necessary to set up 2 different sets of wells for measuring the total choline and the free choline in this assay.

Therefore, to m...

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Thank you for contacting us.

We have not tested this kit with rat samples. However this kit should work as it does not contain any human specific antibody. The species is irrelevant for this product.

It will also be a good idea to inc...

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Thank you for contacting us.

All of the information and images 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. Unfortunatel...

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Thank you very much for contacting Abcam. Your details has been forwarded to me by my colleague Peter.

The kit ab65345 have a detection range of 0.01 nmol/well to 5 nmol/ml; expression outside of this range will be undetectablewith the kit. <...

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Thank you for pointing this out. The datasheet is somewhat mis-leading with regards to the 500-fold range that is specified, and also in error with regards to the notation. I apologize for the confusion.

The specified range uses the low valu...

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Tissue culture SN can be used with this assay. Add up to 50 µl of the SN to wells and make up the final volume in all wells to 50 µl with the assay buffer. Continue with the protocol.

Thank you for your email. I am sorry to hear that you have been experiencing problems with this kit.

I have checked the details you have kindly provided; however I would like to ask few questions in order to understand the problem;


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1-10 of 17 Abreviews or Q&A


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