Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Butyrylcholinesterase/BCHE Immunoassay is a 4.5 hour solid phase ELISA designed to measure BCHE levels in cell culture supernates, cell lysates, tissue lysates, serum, plasma, saliva, urine, and human milk. It contains CHO cell-expressed recombinant human BCHE and antibodies raised against the recombinant protein. Results obtained for naturally occurring human BCHE showed linear curves that were parallel to the standard curves obtained using the Quantikine Human BCHE Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human BCHE.

Preparation and Storage

Background:  Butyrylcholinesterase/BCHE

Butyrylcholinesterase (BCHE) is a major acetylcholine hydrolyzing enzyme in the circulation. Although it is present in significant amounts (~3 mg/L) in human plasma, no endogenous physiological substrate has been described for this enzyme. It can degrade a large number of ester-containing compounds in addition to acylcholines. Thus, it is likely to play significant pharmacological and toxicological roles. It is thought to be involved in the pathological process of Alzheimer's disease (AD) by depleting acetylcholine. In contrast to ACHE, it attenuates amyloid fibril formation in vitro. BCHE inhibitors have been used to delay symptoms of AD patients by virtue of their ability to enhance acetylcholine availability. Its involvement in a cholinergic anti-inflammatory pathway connect BCHE and ACHE with a possible marker of low-grade systemic inflammation observed in Type-2 diabetes, obesity, hypertension, coronary heart disease, and AD. BCHE can exist in monomeric and multimeric forms. The expressed recombinant mouse BCHE contains multiple forms that consist of soluble monomers, dimers, and tetramers.

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.