Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF

R&D Systems | Catalog # 5284-CE

R&D Systems
Discontinued Product
5284-CE has been discontinued. View all Carboxylesterase 5/CES5 products.

Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Carboxylesterase 5/CES5 Protein (5284-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

Applications

Enzyme Activity
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Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Carboxylesterase 5/CES5 protein
Met1-His556, with a C-terminal 10-His tag
Accession # NP_766347

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

No results obtained: Gln27 predicted

Predicted Molecular Mass

61 kDa

SDS-PAGE

60 kDa, under reducing conditions

Activity

Measured by its ability to hydrolyze p-nitrophenylacetate.
The specific activity is >15,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5284-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 5/CES5

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.

References

  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.

Entrez Gene IDs

234673 (Mouse)

Gene Symbol

CES2E

UniProt

Product Documents for Recombinant Mouse Carboxylesterase 5/CES5 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 5/CES5 Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF (5284-CE):

Materials
  • 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

FAQs

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