Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

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DBCHE0
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Human Butyrylcholinesterase/BCHE ELISA Standard Curve
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Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Tissue Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (50 uL), Urine (50 uL), Human Milk (50 uL)
Sensitivity
0.06 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Cell Lysates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
Specificity
Natural and recombinant human BCHE.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

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.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Cell Lysates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.25 2.41 5.01 1.26 2.51 5.11
Standard Deviation 0.031 0.044 0.154 0.063 0.114 0.177
CV% 2.5 1.8 3.1 3.1 4.5 3.5

Recovery

The recovery of human BCHE spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 94-105
Cell Lysis Buffer 1 (n=2) 99 96-102
Urine (n=4) 97 93-102

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human BCHE were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Butyrylcholinesterase/BCHE ELISA Linearity

Data Examples

Human Butyrylcholinesterase/BCHE ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Entrez Gene IDs:
590 (Human); 12038 (Mouse)
Alternate Names:
Acylcholine acylhydrolase; BCHE; Butyrylcholine esterase; Butyrylcholinesterase; CHE1; CHE1cholinesterase; Choline esterase II; cholinesterase 1; E1; EC 3.1.1.8; Pseudocholinesterase
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for 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. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Serum Level and Activity of Butylcholinesterase: A Biomarker for Post-Stroke Dementia
    Authors: YC Chen, WH Chou, CP Fang, TH Liu, HH Tsou, Y Wang, YL Liu
    J Clin Med, 2019;8(11):.
    Species: Human
    Sample Types: Plasma
  2. Adeno-associated virus-mediated expression of human butyrylcholinesterase to treat organophosphate poisoning
    Authors: V Gupta, CL Cadieux, D McMenamin, CA Medina-Jas, M Arif, O Ahonkhai, E Wielechows, M Taheri, Y Che, T Goode, MP Limberis, M Li, DM Cerasoli, AP Tretiakova, JM Wilson
    PLoS ONE, 2019;14(11):e0225188.
    Species: Transgenic Mouse
    Sample Types: Serum

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