Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF

Catalog #: 5284-CE Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
5284-CE-010

R&D Systems Recombinant Proteins and Enzymes

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

Recombinant Mouse Carboxylesterase 5/CES5 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 >15,000 pmol/min/μg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Carboxylesterase 5/CES5 protein
Met1-His556, with a C-terminal 10-His tag
Accession # NP_766347

Accession #

NP_766347

N-terminal Sequence
Analysis

No results obtained: Gln27 predicted

Predicted Molecular Mass

61 kDa

SDS-PAGE

60 kDa, under reducing conditions

Product Datasheets

5284-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.

5284-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: 50 mM Tris, pH 7.5
Recombinant Mouse Carboxylesterase 5/CES5 (rmCES5) (Catalog # 5284-CE)
Substrate: 4-Nitrophenyl acetate (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 rmCES5 to 0.2 ng/µL in Assay Buffer.
Dilute Substrate to 2 mM in deionized water.
Load 50 µL of 0.2 ng/µL rmCES5 into a plate, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
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:

rmCES5: 0.010 µg
Substrate: 1 mM

Reconstitution Calculator

Background: Carboxylesterase 5/CES5

Carboxylesterase 5 encoded by the CES5 gene is a member of a large family of carboxylesterases that are responsible for the hydrolysis of ester and amide bonds (1, 2). They have broad substrate specificity ranging from small molecule esters such as phenylester to long-chain fatty acid esters and thioesters. Carboxylesterases play a major role as determinants of pharmacokinetic behavior for most therapeutic agents containing an ester, participating in the detoxification of drugs such as cocaine and heroin in serum and liver. They can also detoxify organophosphate and carbamate analogues used in agrochemicals or chemical nerve agents, such as malathion, sarin, tabun, and VX. Carboxylesterases can also perform transesterification, a reaction important for cholesterol homeostasis. Carboxylesterase deficiency may be associated with non-Hodgkin lymphoma or B-cell lymphocytic leukemia. CES5 shares the serine hydrolase fold observed in other esterases (3). CES5 possesses an endoplasmic reticulum retention signal (HKEL) at its C-terminus. The expressed recombinant mouse CES5 lacks this signal, resulting in its secretion.

References