Recombinant Mouse Carboxylesterase 1/CES1 Protein, CF

R&D Systems | Catalog # 7929-CE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Carboxylesterase 1/CES1 Protein (7929-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

Q8VCC2

Applications

Enzyme Activity
View all Carboxylesterase 1/CES1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Carboxylesterase 1/CES1 protein
His19-Leu565, with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His19

Predicted Molecular Mass

62 kDa

SDS-PAGE

55-60 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze p-nitrophenylacetate.
The specific activity is >330 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7929-CE
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Carboxylesterase 1/CES1

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). Ces1G is a rat and mouse specific protein that is expressed predominantly in liver, but also in kidney and lung (4).

References

  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.
  4. Ellinghaus P. et al. (1998) Biochim. Biophys. Acta. 1397:175.

Alternate Names

ACAT, CES1, Egasyn, HMSE1, REH, SES1, TGH

Entrez Gene IDs

1066 (Human); 12623 (Mouse); 29225 (Rat)

Gene Symbol

CES1

UniProt

Q8VCC2

Additional Carboxylesterase 1/CES1 Products

Product Documents for Recombinant Mouse Carboxylesterase 1/CES1 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 Mouse Carboxylesterase 1/CES1 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

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Protocols

View specific protocols for Recombinant Mouse Carboxylesterase 1/CES1 Protein, CF (7929-CE):

Materials
  • Assay Buffer: 50 mM Tris, 150 mM NaCl, pH 7.5
  • Reading Buffer: 50 mM Tris, pH 8.0
  • Recombinant Mouse Carboxylesterase 1/CES1 (rmCES1) (Catalog # 7929-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 rmCES1 to 12 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load 50 µL of 12 ng/µL rmCES1 into a clear microplate in triplicate and start the reaction by adding 50 µL of 2 mM 4-NPA. Include a Substrate 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 well (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:
  • rmCES1:  0.6 µg
  • Substrate:  0.5 mM

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