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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.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

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Images

Functional Studies - Choline/Acetylcholine Assay Kit (AB65345), expandable thumbnail
  • Functional Studies - Choline/Acetylcholine Assay Kit (AB65345), expandable thumbnail
  • Functional Studies - Choline/Acetylcholine Assay Kit (AB65345), expandable thumbnail
  • Functional Studies - Choline/Acetylcholine Assay Kit (AB65345), expandable thumbnail
  • Functional Studies - Choline/Acetylcholine Assay Kit (AB65345), expandable thumbnail

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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

Reactive species

Mammals

Range

0.01 - 5 nmol/well

Assay time

40m

Sensitivity

> 0.01 nmol/well

What's included?

2000 Test
Components
Acetylcholinesterase Enzyme
20 x 1 Vial
Assay Buffer VI
20 x 25 mL
Choline Standard
20 x 1 Vial
Enzyme Mix IV
20 x 1 Vial
OxiRed Probe
20 x 200 µL

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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.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

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

Reactive species

Mammals

Range

0.01 - 5 nmol/well

Assay time

40m

Assay Platform

Microplate reader

Sensitivity

> 0.01 nmol/well

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

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 .

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Associated diseases and disorders

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.

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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5 product images

  • Functional Studies - Choline/Acetylcholine Assay Kit (ab65345), expandable thumbnail
    Image from Kim J.H et al., PLoS One 8(2), Fig 4. doi: 10.1371/journal.pone.0056736. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Choline/Acetylcholine Assay Kit (ab65345)

    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).

  • Functional Studies - Choline/Acetylcholine Assay Kit (ab65345), expandable thumbnail

    Functional Studies - Choline/Acetylcholine Assay Kit (ab65345)

  • Functional Studies - Choline/Acetylcholine Assay Kit (ab65345), expandable thumbnail

    Functional Studies - Choline/Acetylcholine Assay Kit (ab65345)

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

  • Functional Studies - Choline/Acetylcholine Assay Kit (ab65345), expandable thumbnail

    Functional Studies - Choline/Acetylcholine Assay Kit (ab65345)

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

  • Functional Studies - Choline/Acetylcholine Assay Kit (ab65345), expandable thumbnail

    Functional Studies - Choline/Acetylcholine Assay Kit (ab65345)

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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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