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Acetylcholinesterase Quantification

Acetylcholinesterase (AChE) is one of the most crucial enzymes for nerve response and function. Through its hydrolytic activity, AChE degrades the neurotransmitter acetylcholine (ACh) into choline and acetic acid. AChE is mainly found at neuromuscular junctions and cholinergic synapses in the central nervous system, where its activity serves to terminate the synaptic transmission. It is also found on the red blood cell membranes, where it constitutes the Yt blood group antigen. AChE exists in multiple molecular forms, which possess similar catalytic properties but differ in their oligomeric assembly and mode of attachment to the surface of cells. In therapeutics, AChE inhibitors are among the key drugs approved for Alzheimer's disease (AD) and myasthenia gravis.

Amplite® Colorimetric Acetylcholinesterase Assay Kit (Cat No. 11400) uses DTNB to quantify the amount of thiocholine produced via the hydrolysis of acetylthiocholine by AChE in blood, in cell extracts, and other solutions. The absorption intensity of the DTNB adduct is used to measure the amount of thiocholine formed, which is proportional to the AChE activity.

Amplite® Fluorimetric Acetylcholinesterase Assay Kits provide one of the most sensitive methods for detecting AChE activity or screening AChE inhibitors in the red fluorescence window. Kit 11402 uses Amplite® Red to quantify the amount of choline produced from the hydrolysis of acetylcholine by AChE through choline oxidase-mediated enzyme coupling reactions. The fluorescence intensity of Amplite® Red is used to measure the amount of choline formed, which is proportional to the AChE activity. Kit 11402 can be used for monitoring and quantifying the AChE activity in blood, cell extracts, or other solutions. The kit is an optimized 'mix and read' assay that provides a simple one-step fluorimetric assay to detect as little as 0.01 mU AChE in a 100 µL assay volume (0.1 mU/mL). Its signal can be easily read at Ex/Em = ~540/590 nm.

Amplite® Fluorimetric Acetylcholinesterase Assay Kit (Cat No. 11401) uses our outstanding Thiolite™ Green to quantify the thiocholine produced from the hydrolysis of acetylthiocholine by AChE. Thiolite™ Green is non-fluorescent until reacted with a thiol group. It has spectral properties similar to fluorescein, making this assay compatible with almost all fluorescence instruments. The fluorescence intensity of Thiolite™ Green is used to measure AChE activity. Compared to the existing thiol probes (e.g., mBBr and bBBr), Thiolite™ Green is much more sensitive. This Amplite® Fluorimetric Acetylcholinesterase Assay Kit provides an ultrasensitive fluorimetric one-step assay to detect as little as 0.01mU AChE in a 100 µL assay volume (0.1 mU/mL). Its signal can be easily read using a fluorescence microplate reader at Ex/Em = 490/525 nm.


Left: Acetylcholinesterase dose responses were measured in a 96-well clear plate with Amplite® Colorimetric Acetylcholinesterase Assay Kit. Center: Acetylcholinesterase dose responses were measured in a 96-well black solid plate with Amplite® Fluorimetric Acetylcholinesterase Assay Kit. Right: Acetylcholinesterase dose responses were measured in a 96-well black solid plate with Amplite® Fluorimetric Acetylcholinesterase Assay Kit (Cat No. 11401).

 

Table 1. Acetylcholinesterase Assay Kits

Cat No.
Product Name
Abs (nm)
Em (nm)
Unit Size
40007Amplite® Choline Quantitation Kit571585200 Tests
11403Amplite® Fluorimetric Acetylcholine Assay Kit *Red Fluorescence*571585200 Tests
11400Amplite® Colorimetric Acetylcholinesterase Assay Kit410 200 Tests
11401Amplite® Fluorimetric Acetylcholinesterase Assay Kit *Green Fluorescence*510524200 Tests
11402Amplite® Fluorimetric Acetylcholinesterase Assay Kit *Red Fluorescence*571584200 Tests