Description
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Urine, Plasma, Cell culture extracts, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids,
Assay type:Quantitative,
Sensitivity:> 0.01 nmol/well,
Range:0.01 - 5 nmol/well,
Assay time:40m,
Assay Platform:Microplate reader
Product details:
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
If you need to purchase additional Choline Probe for this product, please see .
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Choline also known by its alternate name trimethylammonium ethanol is a water-soluble essential nutrient. It serves as a precursor for the neurotransmitter acetylcholine which is critical for numerous neural functions. Acetylcholine a small molecule with a molecular mass of 146.21 g/mol is synthesized in nerve terminals from choline and acetyl-CoA by the enzyme choline acetyltransferase. This target is widely expressed across both the central and peripheral nervous systems and its availability and uptake in nerve cells are important for maintaining optimal levels of acetylcholine.
Biological function summary
Choline and its derivative acetylcholine play important roles in cell membrane integrity and neurotransmission. Acetylcholine a constituent of the cholinergic system functions as a neurotransmitter involved in muscle activation cognitive processes and modulation of arousal. It does not typically form part of a larger complex but works in synaptic clefts to transmit signals between neurons. The presence of acetylcholine depends on the availability of choline the substrate used for its synthesis.
Pathways
Choline and acetylcholine are integral to the cholinergic signaling pathway. Acetylcholine interacts with muscarinic and nicotinic receptors to transmit neural signals. These receptors are vital parts of the pathway that modulates parasympathetic nervous system responses. Choline kinase involved in phosphorylating choline to phosphocholine links the choline metabolism pathway to cell signaling and fatty acid biosynthesis.
Choline and acetylcholine are associated with conditions like Alzheimer's disease and myasthenia gravis. Alzheimer's disease involves decreased levels of acetylcholine due to cholinergic neuron degeneration impacting cognitive functions. In myasthenia gravis an autoimmune disorder antibodies target acetylcholine receptors leading to muscle weakness. Both conditions highlight the importance of acetylcholine's relation to choline acetyltransferase as this enzyme's activity is pivotal for maintaining acetylcholine levels essential for normal nervous system function.
