Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric/Fluorometric
- Platform: Microplate reader
- Assay time: 40 min
- Sample type: Cell culture extracts, Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
- Sensitivity: 0.01 nmol/well
Product nameCholine/Acetylcholine Assay Kit
Sample typeCell culture supernatant, Urine, Serum, Plasma, Cell culture extracts, Other biological fluids, Tissue Extracts
Sensitivity> 0.01 nmol/well
Range0.01 nmol/well - 5 nmol/well
Assay time0h 40m
Species reactivityReacts with: Mammals, Other species
Choline/Acetylcholine Assay Kit (ab65345) provides a simple and sensitive means for quantifying Choline and Acetylcholine by either a colorimetric or fluorometric method in samples such as blood, cells, culture media, fermentation media, etc.
In the choline / acetylcholine assay protocol, 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 to measure total choline (choline + acetyl choline). The amount of acetyl choline is calculated by subtracting choline from total choline.
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.
Choline / acetylcholine assay protocol summary:
- add samples and standards to wells
- add choline reaction mix with, or without, acetylcholinesterase, and incubate for 30 min at room temp
- analyze with microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K615 Choline/Acetylcholine Quantification Colorimetric/Fluorometric Kit. K615-100 is the same size as the 100 test size of ab65345.
If you need to purchase additional Choline Probe for this product, please see ab139505.
Storage instructionsStore at -20°C. Please refer to protocols.
Storage bufferPreservative: None
Components Identifier 100 tests 2000 tests Acetylcholinesterase Blue 1 vial 20 vials Choline Assay Buffer WM 1 x 25ml 20 x 25ml Choline Enzyme Mix Green 1 vial 20 vials Choline Probe (in DMSO) Red 1 x 200µl 20 x 200µl Choline Standard (5 µmol) Yellow 1 vial 20 vials
RelevanceCholine and acetylcholine (often abbreviated ACh) play important roles in many biological processes. Choline is a natural amine, and is generally found in phosphatidylcholine and sphingomyelin, phopholipids abundant in cell membranes. Choline is also the precursor molecule for the neurotransmitter acetylcholine which is involved in many functions both in the peripheral nervous system (PNS) and in the central nervous system (CNS). In the PNS, ACh activates muscles and is a major neurotransmitter in the autonomic nervous system. In the CNS, ACh and the associated neurons form the cholinergic system, which tends to cause anti-excitatory actions.
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
ab65345 has been referenced in 28 publications.
- Qian X et al. Dopamine Pathway Mediated by DRD5 Facilitates Tumor Growth via Enhancing Warburg Effect in Esophageal Cancer. Front Oncol 11:655861 (2021). PubMed: 33898321
- Cortés-Gómez MÁ et al. Tau phosphorylation by glycogen synthase kinase 3ß modulates enzyme acetylcholinesterase expression. J Neurochem 157:2091-2105 (2021). PubMed: 32955735
- Daniele S et al. The Nootropic Drug ?-Glyceryl-Phosphoryl-Ethanolamine Exerts Neuroprotective Effects in Human Hippocampal Cells. Int J Mol Sci 21:N/A (2020). PubMed: 32023864
- Liu EYL et al. Interacting with a 7 nAChR is a new mechanism for AChE to enhance the inflammatory response in macrophages. Acta Pharm Sin B 10:1926-1942 (2020). PubMed: 33163344
- London E et al. Loss of habenular Prkar2a reduces hedonic eating and increases exercise motivation. JCI Insight 5:N/A (2020). PubMed: 33141766