Recombinant Mouse Butyrylcholinesterase/BCHE Protein, CF

Catalog #: 5527-CE Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
5527-CE-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Butyrylcholinesterase/BCHE Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to cleave Butyrylthiocholine. The specific activity is >75 nmol/min/μg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Butyrylcholinesterase/BCHE protein
His28-Leu603, with a C-terminal 6-His tag

Accession #

AAH99977

N-terminal Sequence
Analysis

His28

Predicted Molecular Mass

66 kDa

SDS-PAGE

86 kDa, reducing conditions

Product Datasheets

5527-CE

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

5527-CE

Formulation	Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials

Assay Buffer: 100 mM Sodium Phosphate, pH 7.5
Recombinant Mouse Butyrylcholinesterase/BCHE (rmBCHE) (Catalog # 5527-CE)
Substrate: S-Butyrylthiocholine chloride (BTC) (Sigma Catalog # B-3128), 20 mM stock in DMSO
5,5’-dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma Catalog # D-8130), 10 mM stock in DMSO
96 well Clear Plate (Costar, Catalog # 92592)
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

Dilute rmBCHE to 0.01 µg/mL in Assay Buffer.
Dilute BTC to 10 mM in deionized water.
Combine equal amounts of diluted BTC and DTNB. Dilute substrate mixture to a final concentration 200 µM BTC and DTNB in deionized water.
Load into plate 50 µL of 0.01 µg/mL rmBCHE and start the reaction by adding 50 µL of the BTC/DTNB mixture to the wells. As a Substrate Blank, load 50 µL of Assay Buffer and 50 µL of the BTC/DTNB mixture.
Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
Calculate specific activity:

Specific Activity (nmol/min/µg) =	Adjusted V_max* (OD/min) x well volume (L) x 10⁹ nmol/M
	ext. coeff (M^-1cm^-1) x path corr.* (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Using the extinction coefficient 13260 M^-1cm^-1
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:

rmBCHE: 0.0005 µg
DTNB and BTC: 100 µM

Reconstitution Calculator

Background: Butyrylcholinesterase/BCHE

Butyrylcholinesterase (BCHE) is a major acetylcholine hydrolyzing enzyme in the circulation (1). 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 (2). BCHE inhibitors have been used to delay symptoms of AD patients by virtue of their ability to enhance acetylcholine availability (3). 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 (4). BCHE can exist in monomeric and multimeric forms (1). The expressed recombinant mouse BCHE contains multiple forms that consist of soluble monomers, dimers, and tetramers.

References

Darvesh, S. et al. (2003) Nat. Rev. Neuroscience 4:131.
Diamant, S. et al. (2006) Proc. Natl. Acad. Sci. USA 103:8628.
Campbell, V. A. and Gowran, A. (2007) Br. J. Pharm. 152:655.
Das, U. N. (2007) Med Sci Monit. 13:RA214.

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