Recombinant Mouse Carboxylesterase 3/CES3 Protein, CF

Catalog #: 5428-CE Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
5428-CE-010

R&D Systems Recombinant Proteins and Enzymes

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All Carboxylesterase 3/CES3 Proteins and Enzymes

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

Recombinant Mouse Carboxylesterase 3/CES3 Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain

Endotoxin Level

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

Activity

Measured by its ability to hydrolyze p-nitrophenylacetate. The specific activity is >11,000 pmol/min/μg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Carboxylesterase 3/CES3 protein
Tyr19-Glu561, with a C-terminal 10-His tag

Accession #

Q8VCT4

N-terminal Sequence
Analysis

Tyr19

Predicted Molecular Mass

61 kDa

SDS-PAGE

58 kDa, reducing conditions

Product Datasheets

5428-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.

5428-CE

Formulation	Supplied as a 0.2 μm filtered solution in MES 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: 50 mM Tris, pH 7.5
Recombinant Mouse Carboxylesterase 3/CES3 (rmCES3) (Catalog # 5428-CE)
Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N-8130), 100 mM stock in Acetone
96-well Clear Plate (Costar, Catalog # 92592)
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

Dilute rmCES3 to 0.2 ng/µL in assay buffer.
Dilute substrate 4-NPA to 2 mM in deionized water.
Load in a clear microplate 50 µL of 0.2 ng/µL rmCES3, and start the reaction by adding 50 µL of 2 mM 4-NPA. Include a blank consisting of 50 µL of assay buffer and 50 µL 2 mM 4-NPA.
Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (OD/min) x Conversion Factor** (pmol/OD)
	amount of enzyme (µg)

*Adjusted for Substrate Blank

**Derived using calibration standard 4-Nitrophenol (Sigma, Catalog # 241326).

Per Well:

rmCES3: 0.010 µg
Substrate: 1 mM

Reconstitution Calculator

Background: Carboxylesterase 3/CES3

Carboxylesterase 3 is a member of a large family of carboxylesterases that are responsible for the hydrolysis of ester and amide bonds (1, 2). These carboxylesterases are widely distributed in mammalian tissues and have broad substrate specificity ranging from small molecule esters such as phenylester to long‑chain fatty acid esters and thioesters. Because many ester-containing drugs require the ester linkage to improve the bioavailability of therapeutic agents, CESs play a major role in drug metabolism and activation. CES3 is also known as triacylglycerol hydrolase for its lipolytic activity. It has been reported to be the major lipase in white adipose tissue (3).

References