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Recombinant Human Carboxylesterase 2/CES2 Protein, CF

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Recombinant Human CES-2/His (Catalog # 5657-CE) has a molecular weight (MW) of 64.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Recombinant Human CES-2/His (Catalog # 5657-CE) has a molecular weight (MW) of 64.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer.  MW may differ from predicted MW due to post-translational ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Carboxylesterase 2/CES2 Protein, CF Summary

Additional Information
Analyzed by SEC-MALS
Details of Functionality
Measured by its ability to hydrolyze p-nitrophenylacetate. The specific activity is >30,000 pmol/min/μg, measured under the described conditions. 
Source
Mouse myeloma cell line, NS0-derived human Carboxylesterase 2/CES2 protein
Gln27-Leu559, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
No results obtained: Gln27 predicted
Structure / Form
Monomer
Protein/Peptide Type
Recombinant Enzymes
Gene
CES2
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
60 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
63 kDa, reducing conditions
Publications
Read Publications using
5657-CE in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate, NaCl and Glycerol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Assay Procedure
  • Assay Buffer: 50 mM Tris, pH 7.5
  • Reading Buffer: 50 mM Tris, pH 8.0
  • Recombinant Human Carboxylesterase 2/CES2 (rhCES2) (Catalog # 5657-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
  1. Dilute rhCES2 to 0.1 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in deionized water.
  3. Load into a clear microplate 50 µL of 0.1 ng/µL rhCES2 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 of 2 mM 4-NPA.
  4. Incubate for 10 minutes at room temperature.
  5. Add 100 µL Reading Buffer to each reaction (this will not stop the reaction).
  6. Read immediately at a wavelength of 410 nm (bottom read) in endpoint mode.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)

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

Per Well:
  • rhCES2: 0.005 µg
  • Substrate: 0.5 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Carboxylesterase 2/CES2 Protein, CF

  • carboxylesterase 2 (intestine, liver)
  • Carboxylesterase 2
  • carboxylesterase 2EC 3.1.1.1
  • CE-2
  • CE-2PCE-2
  • CES2
  • CES2A1
  • EC 3.1.1
  • EC 3.1.1.84
  • hCE-2
  • iCE
  • intestinal carboxylesterase; liver carboxylesterase-2
  • PCE-2

Background

Carboxylesterase 2 is a member of a serine esterase family composed of enzymes which hydrolyze ester and amide bonds (1, 2). The members in this family share the serine hydrolase fold observed in other esterases (3). They have broad substrate specificity from small molecule esters such as phenylester to long-chain fatty acid esters and thioesters. They play a major role in the pharmacokinetics of most therapeutic agents containing an ester. By de-esterification, they can activate or inactivate the agents. They also participate in the detoxification of drugs such as cocaine and heroin in serum and liver. In addition to narcotics, they can also detoxify organophosphate and carbamate analogues used in agrochemicals or chemical nerve agents, such as malathion, sarin, tabun, and VX. In addition to the hydrolytic activity, they can perform transesterification. This reaction is important for cholesterol homeostasis. Three major human CESs have been identified (4). CES1 is highly expressed in liver. CES2 is present in the small intestine, colon, kidney, liver, heart, brain, and testis. CES3 is brain-specific. Carboxylesterase deficiency may be associated with non-Hodgkin lymphoma or B-cell lymphocytic leukemia.

  1. Redinbo, M. R. and P.M. Potter. (2005) Drug Discovery Today, 10:313.
  2. Satoh, T. and M. Hosokawa. (2006) Chem.-Biol. Interactions, 162:195.
  3. Fleming, C. D. et al. (2007) Biochemistry 46:5603.
  4. Imai, T. (2006) Drug Metab. Pharmacokinet. 21:173.

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Recombinant Human
Carboxylesterase 2/CE ...
5657-CE
Recombinant Human Carboxylesterase 2/CES2 Protein, CF
Species: Hu
Applications: Enzyme Activity

Publications for Carboxylesterase 2/CES2 (5657-CE)(3)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: Bioassay, Enzyme Assay.


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Bioinformatics

Gene Symbol CES2
Uniprot