Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

R&D Systems | Catalog # DBCHE0

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Tissue Lysates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Saliva (50 µL), Urine (50 µL), Human Milk (50 µL)

Sensitivity

0.06 ng/mL

Assay Range

0.156-10 ng/mL (Cell Culture Supernates, Cell Lysates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
View all Butyrylcholinesterase/BCHE ELISA Kits »

Product Summary for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Tissue Lysates, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 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 for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

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

Scientific Data Images for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

Human Butyrylcholinesterase/BCHE ELISA Standard Curve

Human Butyrylcholinesterase/BCHE ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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.

Alternate Names

Acylcholine acylhydrolase, BCHE, Butyrylcholine esterase, CHE1, Pseudocholinesterase

Entrez Gene IDs

590 (Human); 12038 (Mouse)

Gene Symbol

BCHE

Additional Butyrylcholinesterase/BCHE Products

Product Documents for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit

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Protocols

View specific protocols for Human Butyrylcholinesterase/BCHE Quantikine ELISA Kit (DBCHE0):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  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 on a horizontal orbital microplate shaker.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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