Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF
Recombinant Mouse Carboxylesterase 5/CES5 Protein, CF Summary
Product Specifications
Met1-His556, with a C-terminal 10-His tag
Accession # NP_766347
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
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.
|
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
- Dilute rmCES5 to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in deionized water.
- 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.
- Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
- 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).
- rmCES5: 0.010 µg
- Substrate: 1 mM
Reconstitution Calculator
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.
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