Acetylcholinesterase (AChE) Fluorescent Activity Kit

Documentation

Specifications

• Assay Type Activity Assay
• Sample Types Serum, Plasma (EDTA and Heparin), RBC Ghosts, Extracted Tissue Sample, Erythrocyte Membranes
• Sensitivity 0.228 mU/mL
• Species Species Independent
• Assay Duration 20 Minutes
• Samples/Plate 42 in Duplicate
• Readout Fluorescent, 510 nm Emission / 390 nm Excitation
• Standard Curve
• Description

  Assay Principle: 

  The Acetylcholinesterase (AChE) Fluorescent Activity Kit quantitatively measures AChE activity in serum, plasma (EDTA and Heparin), RBC ghosts, and extracted tissue samples. The Acetylcholinesterase (AChE) Fluorescent Activity Kit is an Activity Assay with a run time of 20 minutes. Please read the complete kit insert for more information before performing this assay.

  Use our provided Acetylcholinesterase Standard to generate a standard curve for the assay. Pipette the standards or diluted samples into the black microtiter plate. Add the Assay Buffer and Reaction Mix to each well, tapping the plate to ensure sufficient mixing of reagents. Then incubate the plate at room temperature for 20 minutes. The fluorescent-generating reaction occurs when the ThioStar® reagent binds to the thiol generated between the AChE substrate and the AChE present in the samples.

  After the 20-minute incubation, use a plate reader to detect and record the generated fluorescent signal at 510nm. Use the intensity and the standard curve to calculate the AChE activity in the samples.

  Background:

  Acetylcholinesterase (AChE) is an enzyme that breaks down the key neurotransmitter Acetylcholine (ACh). AChE appears critical to both the development and function of the nervous system. Acetylcholine is an essential neurotransmitter for cholinergic neurons in the central and peripheral nervous systems. Cholinergic neurons are essential in regulating the circadian rhythm and modulating memory processes. Impairment of cholinergic neurotransmission is well-established in Alzheimer’s disease (AD). AChE inhibitors are established therapeutics that inhibit ACh breakdown and prolong cholinergic neuronal firing. These inhibitors are the most common form of AD treatment. The use of AChE inhibitors as therapeutic agents and pesticides has spurred detailed investigations of cholinesterases since their identification.