Human Acetylcholinesterase/ACHE DuoSet ELISA Summary

Product Summary

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Acetylcholinesterase (ACHE). The suggested diluent is suitable for the analysis of most cell culture supernate, cell lysate, serum, and plasma samples. Diluents for complex matrices, such as cell lysates, serum, and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Background: Acetylcholinesterase/ACHE

The classical role of ACHE is to terminate cholinergic neurotransmission by hydrolysis of acetylcholine (ACH). ACHE is thought to be involved in the pathology of Alzheimer's disease (AD) by accelerating the assembly of Abeta peptides into fibrillar species through forming complexes with Abeta via the peripheral anionic site on ACHE. ACHE inhibitors have been used to delay symptoms of AD patients by virtue of their ability to enhance ACH availability, as well as reduce amyloidogenesis and subsequent neurotoxicity. Its involvement in the cholinergic anti-inflammatory pathway connects ACHE with a possible marker of low-grade systemic inflammation in obesity, hypertension, coronary heart disease, and AD. Alternative splicing produces three isoforms: an amphipathic form that exists as both monomeric and mutimeric forms, a soluble-monomeric form lacking the cysteine residue near the C-terminus, and a GPI-anchored dimeric form found in the membranes of erythrocytes. The recombinant mouse ACHE (rmACHE) was expressed as the amphipathic form that consists of soluble monomer and mutimeric forms.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1、Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2、Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3、Add 50 μL of Assay Diluent to each well.

4、Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.