Recombinant Human Carboxylesterase 2/CES2 Protein, CF

R&D Systems | Catalog # 5657-CE

Analyzed by SEC-MALS
R&D Systems
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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Carboxylesterase 2/CES2 Protein (5657-CE)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

Structure / Form

Monomer

Applications

Enzyme Activity
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Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Carboxylesterase 2/CES2 protein
Gln27-Leu559, with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

No results obtained: Gln27 predicted

Predicted Molecular Mass

60 kDa

SDS-PAGE

63 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze p-nitrophenylacetate.
The specific activity is >30,000 pmol/min/μg, measured under the described conditions. 

Scientific Data Images for Recombinant Human Carboxylesterase 2/CES2 Protein, CF

Recombinant Human Carboxylesterase 2/CES2 Protein SEC-MALS.

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).

Formulation, Preparation, and Storage

5657-CE
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate, NaCl and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Carboxylesterase 2/CES2

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.

References

  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.

Alternate Names

CE-2, CES2, PCE-2

Entrez Gene IDs

8824 (Human); 234671 (Mouse); 171118 (Rat)

Gene Symbol

CES2

UniProt

Additional Carboxylesterase 2/CES2 Products

Product Documents for Recombinant Human Carboxylesterase 2/CES2 Protein, CF

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 Recombinant Human Carboxylesterase 2/CES2 Protein, CF

For research use only

Citations for Recombinant Human Carboxylesterase 2/CES2 Protein, CF

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Protocols

View specific protocols for Recombinant Human Carboxylesterase 2/CES2 Protein, CF (5657-CE):

Materials
  • 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

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