EnzyChrom™ Acetylcholine Assay Kit

SKU:BHT15600110
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    Overview
    Click light‑blue chips for details
    EnzyChrom Acetylcholine Assay Kit is designed for quantitative determination of acetylcholine and evaluation of drug effects on acetylcholine metabolism. It uses OD570 nm, or FL530/585 nm readout; suited to serum, plasma, urine; typical assay time 30 min; detection limit OD, FL: 10, 0.4 µM.
    Detection method Colorimetric (OD 570 nm) or Fluorescent (FL 530/585 nm)
    Sample type Serum, plasma, urine, saliva, milk, tissue, cell culture, et
    Species All
    Procedure 30 min
    Detection limit Colorimetric: 10 µM / Fluorescent: 0.4 µM
    Available Options

    Select the variant that best fits your experiment. Availability and lead time may vary by option.

    • Options: Size: 100 Tests
    • Lead time: varies by selected option; please contact us for current fulfillment timing.
    • Storage: -20°C — Store at -20°C (freezer). Avoid repeated freeze-thaw cycles.
    • Shipping: cold-chain shipment (typically with ice packs).
    • Upon receipt: store at the recommended temperature as soon as possible.
    • Sales terms and conditions: Please review prior to ordering.
    Options selector
    Catalog no. Size
    EACL-100 100 Tests
    Field Specification
    Assay Time
    • 30 min
    Detection Method
    • Colorimetric (OD 570 nm) or Fluorescent (FL 530/585 nm)
    Product Type
    • Assay Kits
    • Enzyme Activity
    Sample Type(s) Serum, plasma, urine, saliva, milk, tissue, cell culture, etc
    Shipping Cold pack (ICE) — Ships on ice (cold pack included). Store immediately upon receipt.
    Species All
    Storage -20°C — Store at -20°C (freezer). Avoid repeated freeze-thaw cycles.

    Overview

    For quantitative determination of acetylcholine and evaluation of drug effects on acetylcholine metabolism. The assay uses OD570nm, or FL530/585nm for signal readout. Compatible sample input includes Serum, plasma, urine, saliva, milk, tissue, cell culture, etc. Typical stated assay timing is 30 min.

    Key elements and design rationale

    • Readout format: OD570nm, or FL530/585nm supports plate-based signal acquisition and consistent comparison across matched samples.
    • Sample compatibility: The stated sample scope includes Serum, plasma, urine, saliva, milk, tissue, cell culture, etc, which is useful when aligning matrix type with calibration and control design.
    • Analytical range context: The supplied specifications include a stated detection limit of OD, FL: 10, 0.4 µM for interpreting low-signal samples.

    Available format information for this listing includes 100 Tests.

    Biological background

    This product is centered on measurement of acetylcholine within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.

    More details

    ACETYLCHOLINE is a neurotransmitter produced in acetylcholinergic neurons. It plays important roles in skeletal muscle movement, regulation of smooth and cardiac muscles, as well as in learning, memory, and mood. BioAssay Systems method provides a simple, direct, and high-throughput assay for measuring acetylcholine in biological samples. In this assay, acetylcholine is hydrolyzed by acetylcholinesterase to choline which is oxidized by choline oxidase to betaine and H2O2. The resulting H2O2reacts with a specific dye to form a pink-colored product. The color intensity at 570nm or fluorescence intensity (530/585 nm) is directly proportional to the acetylcholine concentration in the sample.

    Detection method

    Colorimetric (OD 570 nm) or Fluorescent (FL 530/585 nm).

    Detection limit and analytical sensitivity

    Reported detection limit(s): Colorimetric: 10 µM / Fluorescent: 0.4 µM. Additional source wording: OD, FL: 10, 0.4 µM.

    Procedures and timing

    Stated procedure or timing information: 30 min.

    Research relevance and current trends

    • Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
    • The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
    • Short assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.

    Common research applications

    • Quantify acetylcholine in serum, plasma, urine by OD570 nm, or FL530/585 nm readout.
    • Compare treatment or phenotype groups using matched serum, plasma, urine handling.
    • Monitor time-course or pre/post changes in serum, plasma, urine across study conditions.

    Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.

    Notes for experimental interpretation

    • Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
    • Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
    Can the acetylcholine level in human serum be measured with this kit?

    We have not tested this assay with blood samples. In the literature search I found the following reference:

    Plasma levels in young women: 456.1 +/- 53.1 (SEM) pg/mL which equals 3.1 nM. (Kawashima et al. Neuroscience Letters 80:(3), 339-342; 1987.) If this is indeed a representative study, it would mean that our assay is not sensitive enough to detect acetylcholine in plasma or serum samples. Furthermore, plasma levels of choline are much higher (~10 &microM) and will thus have to be removed before assaying.

    What is the principle of this assay?

    The assay is based on the enzymatic degradation of acetylcholine to choline and acetate and the subsequent detection of choline.

    Can this assay measure acetylcholine in brain tissue?

    We have not tested acetylcholine assay specifically with brain tissue samples, but believe it should work. Typical concentrations of choline and acetylcholine in brain tissue are 10-70 nmoles/g. So if the tissue sample is suspended in ice-cold PBS at 500 mg/mL, we should expect 5-35 µM choline or acetylcholine in the supernatant.

    Suggested protocol: weigh brain tissue (approx. 50 mg) and add 100 µL ice-cold PBS, lyze tissue by homogenization or sonication. Centrifuge at 14,000 rpm for 10 min at 4°C. Carefully transfer the supernatant to a clean tube and perform the assays for choline and acetylcholine.

    Due to the low concentration range, the fluorescence assay procedure is recommended.

    For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.

    Total isoflavones from soybean and tempeh reversed scopolamine-induced amnesia, improved cholinergic activities and reduced neuroinflammation in brain

    Ahmad, A., Ramasamy, K., Jaafar, S. M., Majeed, A. B. A., & Mani, V. (2014). Total isoflavones from soybean and tempeh reversed scopolamine-induced amnesia, improved cholinergic activities and reduced neuroinflammation in brain. Food and Chemical Toxicology 65: 120-128. Assay: Acetylcholine in rat brain.

    Withania somnifera root extract ameliorates hypobaric hypoxia induced memory impairment in rats

    Baitharu, I., Jain, V., Deep, S. N., Hota, K. B., Hota, S. K., Prasad, D., & Ilavazhagan, G. (2013). Withania somnifera root extract ameliorates hypobaric hypoxia induced memory impairment in rats. Journal of ethnopharmacology 145(2): 431-441. Assay: Acetylcholine in rat hippocampus.

    Acute exposure to DE-71: Effects on locomotor behavior and developmental neurotoxicity in zebrafish larvae

    Chen L et al (2012). Acute exposure to DE-71: Effects on locomotor behavior and developmental neurotoxicity in zebrafish larvae. Environ Toxicol Chem. 31(10):2338-44. Assay: Acetylcholine in fish.

    Embryonic exposure to benzo(a)pyrene influences neural development and function in rockfish (Sebastiscus marmoratus)

    He C, et al (2012). Embryonic exposure to benzo(a)pyrene influences neural development and function in rockfish (Sebastiscus marmoratus). Neurotoxicology 33(4):758-62. Assay: Acetylcholine in fish.

    Exposure of Sebastiscus marmoratus embryos to pyrene results in neurodevelopmental defects and disturbs related mechanisms

    He C, et al (2012). Exposure of Sebastiscus marmoratus embryos to pyrene results in neurodevelopmental defects and disturbs related mechanisms. Aquat Toxicol 15(116-117):109-15. Assay: Acetylcholine in fish.

    Effects of the Total Alkaloidal Extract of Murraya koenigii Leaf on Oxidative Stress and Cholinergic Transmission in Aged Mice

    Mani V, et al (2012). Effects of the Total Alkaloidal Extract of Murraya koenigii Leaf on Oxidative Stress and Cholinergic Transmission in Aged Mice. Phytother Res. 27(1):46-53. Assay: Acetylcholine in mouse brain.

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