EnzyChrom™ Acetylcholine Assay Kit

-20 °C

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 (ca.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 ?M choline or acetylcholine in the supernatant.

Suggested protocol: weigh brain tissue (approx. 50 mg) and add 100 uL ice-cold PBS, lyze tissue by homogenization or sonication. Centrifuge at 14, 000 rpm for 10 min at 4C. 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 more detailed product information and questions, please feel free to send an inquiry to info@hoelzel.de

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