Recombinant Mouse Carboxylesterase 3/CES3 Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse Carboxylesterase 3/CES3 Protein, CF Summary

• Details of Functionality: 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: Tyr19
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Ces1d
• 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: 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: 58 kDa, reducing conditions

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 MES 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 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
  1. Dilute rmCES3 to 0.2 ng/µL in assay buffer.
  2. Dilute substrate 4-NPA to 2 mM in deionized water.
  3. 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.
  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:
  • rmCES3: 0.010 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 Mouse Carboxylesterase 3/CES3 Protein, CF

• carboxylesterase 3 (brain)
• Carboxylesterase 3
• CES3
• EC 3.1.1
• EC 3.1.1.1
• ES31FLJ21736
• esterase 31
• Liver carboxylesterase 31 homolog

Background

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).

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. Soni, K. G. et al. (2004) J. Biol. Chem. 279:40683.

Bioinformatics

• Gene Symbol: Ces1d
• Uniprot:
  • Q8VCT4()