Shelf life (days): 365
Info: Cayman’s Acetylcholine/Choline Fluorometric/Colorimetric Assay Kit provides methods for measuring acetylcholine, choline, and acetylcholine + choline (total) levels in biological samples, such as tissue homogenates, cell culture medium, serum, and plasma. In the assay, the Acetylcholine Converter Enzyme catalyzes the conversion of acetylcholine to choline, which is further converted to betaine aldehyde and hydrogen peroxide (H2O2) by the Choline Detection Enzyme. In the presence of horseradish peroxidase (HRP), H2O2 reacts stoichiometrically with MaxiProbe to produce a fluorescent compound. Background from H2O2 in the sample is determined by omitting the converter and detection enzymes in sample background reactions. To determine choline levels alone, the Acetylcholine Converter Enzyme is excluded from the reaction mixture. The concentration of acetylcholine can be calculated by subtracting choline levels from the total measurement.This assay offers the option to quantify by either fluorescence or absorbance. For fluorescence detection, measurements can be taken using excitation and emission wavelengths of 530 and 590 nm, respectively. Alternatively, the absorbance can be measured at 570 nm. It is at the user’s discretion to choose the mode of measurement (and corresponding standard curve preparation) that best fits their needs. When read fluorometrically, this assay has a range of 0-10 µM and a lower limit of detection (LLOD) of 0.005 µM and 0.04 µM for the choline and total acetylcholine+choline assays, respectively. When read colorimetrically, the range is 0-100 µM with an LLOD of 0.04 µM and 0.07 µM for the choline and total acetylcholine+choline assays, respectively.
Assay Range: 0-10 µM (fluorometric); 0-100 µM (colormetric)
Formula Weight: 0
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