Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF

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

Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF Summary

• Details of Functionality: 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: No results obtained: Gln27 predicted
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Ces2e
• 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: 60 kDa, under 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 Tris 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 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
  1. Dilute rmCES5 to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in deionized water.
  3. 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.
  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:
  • rmCES5: 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 5/CES5 Protein, CF

• Carboxylesterase 5

Background

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.

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.

Bioinformatics

• Gene Symbol: Ces2e
• Uniprot:
  • NP_766347()