Recombinant Human Carboxylesterase 1/CES1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Carboxylesterase 1/CES1 Protein, CF Summary

Details of Functionality
Measured by its ability to hydrolyze p-nitrophenylacetate. The specific activity is >6,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Carboxylesterase 1/CES1 protein
His19-Glu563, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
His19
Protein/Peptide Type
Recombinant Enzymes
Gene
CES1
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

Theoretical MW
61 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
61 kDa, reducing conditions
Publications
Read Publications using
4920-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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM Tris, pH 7.5
  • Recombinant Human Carboxylesterase 1/CES1 (rhCES1) (Catalog # 4920-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 rhCES1 to 0.8 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in deionized water.
  3. In a plate load 50 μL of rhCES1 to wells and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Substrate and 50 µL Assay Buffer.
  4. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (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:
  • rhCES1: 0.040 µg
  • Substrate: 1 mM

Notes

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

Alternate Names for Recombinant Human Carboxylesterase 1/CES1 Protein, CF

  • ACAT
  • Carboxylesterase 1
  • CES1
  • Ces-1
  • Egasyn
  • HMSE1
  • REH
  • SES1
  • Ses-1
  • TGH

Background

Carboxylesterase 1 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. They play a major role as determinants of pharmacokinetic behavior for most therapeutic agents containing an ester. By de-esterification, they can activate or inactivate the agents. They also participate in detoxification of drugs such as cocaine and heroin in serum and liver. The resulting de-esterified metabolites are secreted out in urine. 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, a reaction important for cholesterol homeostasis. Carboxylesterase deficiency may be associated with non-Hodgkin lymphoma or B-cell lymphocytic leukemia. CES-1 shares the serine hydrolase fold observed in other esterases (3). CES-1 possesses an endoplasmic reticulum retention signal (HIEL) at its C-terminus. The purified rhCES-1 lacks this signal, resulting in its secretion.

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

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Publications for Carboxylesterase 1/CES1 (4920-CE)(2)

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Bioinformatics

Gene Symbol CES1
Entrez
Uniprot