Recombinant Mouse Carboxylesterase 31-like/EG13909, CF

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

Recombinant Mouse Carboxylesterase 31-like/EG13909, CF Summary

• Details of Functionality: Measured by its esterase activity. The specific activity is >750 pmol/min/μg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse Carboxylesterase 31-like/EG13909 protein
  Met1-Glu558, with a C-terminal 10-His tag
  Accession # Q8VCU1
• Accession #: Q8VCU1
• N-terminal Sequence: Pro29
• Structure / Form: Monomer
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Ces3b
• 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: 62 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 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
  • Absorbance Buffer: 50 mM Tris, pH 8.0
  • Recombinant Mouse Carboxylesterase 31 like/EG13909 (rmEG13909) (Catalog # 6059-CE)
  • Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rmEG13909 to 2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2.0 mM in Assay Buffer.
  3. Load 50 µL of 2 ng/µL rmEG13909 into plate and start the reaction by adding 50 µL of 2.0 mM Substrate. As a Substrate Blank combine 50 µL Assay Buffer with 50 µL Substrate.
  4. Cover and incubate at room temperature for 60 minutes in the dark.
  5. After incubation add 100 µL Absorbance Buffer and read plate immediately.
  6. Read (top read) absorbance in endpoint mode at 410 nm.
  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 p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).

  Per Well:
  • rmEG13909: 0.1 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 Mouse Carboxylesterase 31-like/EG13909, CF

• Carboxylesterase 31 like
• Carboxylesterase 31-like
• EG13909

Background

EG13909, also known as Es-31, is a carboxylesterase-like enzyme identified in mouse liver (1). It belongs to the type B carboxylesterase/lipase superfamily that hydrolyzes the carboxylic ester bonds. By sequence analysis it is most closely related to mouse CES-3 and is likely to be a microsomal enzyme. It has been suggested that EG13909 is expressed predominantly in male livers and might be regulated by growth hormone (1).

1. Aida, K. et al. (1993) Biochim. Biophys. Acta. 1174:72.

Bioinformatics

• Gene Symbol: Ces3b
• Uniprot:
  • Q8VCU1()