Recombinant Human Carboxylesterase 2/CES2 Protein, CF
Recombinant Human Carboxylesterase 2/CES2 Protein, CF Summary
Product Specifications
Gln27-Leu559, with a C-terminal 10-His tag
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.
5657-CE
| Formulation | Supplied as a 0.2 μm filtered solution in Sodium Acetate, NaCl and Glycerol. |
| Shipping | The product is shipped with dry ice or equivalent. 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
- Reading Buffer: 50 mM Tris, pH 8.0
- Recombinant Human Carboxylesterase 2/CES2 (rhCES2) (Catalog # 5657-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
- Dilute rhCES2 to 0.1 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in deionized water.
- Load into a clear microplate 50 µL of 0.1 ng/µL rhCES2 and start the reaction by adding 50 µL of 2 mM 4-NPA. Include a blank consisting of 50 µL of Assay Buffer and 50 µL of 2 mM 4-NPA.
- Incubate for 10 minutes at room temperature.
- Add 100 µL Reading Buffer to each reaction (this will not stop the reaction).
- Read immediately at a wavelength of 410 nm (bottom read) in endpoint mode.
- 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).
- rhCES2: 0.005 µg
- Substrate: 0.5 mM
Scientific Data
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Recombinant Human CES-2/His (Catalog # 5657-CE) has a molecular weight (MW) of 64.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Reconstitution Calculator
Background: Carboxylesterase 2/CES2
Carboxylesterase 2 is a member of a serine esterase family composed of enzymes which hydrolyze ester and amide bonds (1, 2). The members in this family share the serine hydrolase fold observed in other esterases (3). They have broad substrate specificity from small molecule esters such as phenylester to long-chain fatty acid esters and thioesters. They play a major role in the pharmacokinetics of most therapeutic agents containing an ester. By de-esterification, they can activate or inactivate the agents. They also participate in the detoxification of drugs such as cocaine and heroin in serum and liver. In addition to narcotics, 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. This reaction is important for cholesterol homeostasis. Three major human CESs have been identified (4). CES1 is highly expressed in liver. CES2 is present in the small intestine, colon, kidney, liver, heart, brain, and testis. CES3 is brain-specific. Carboxylesterase deficiency may be associated with non-Hodgkin lymphoma or B-cell lymphocytic leukemia.
- Redinbo, M. R. and P.M. Potter. (2005) Drug Discovery Today, 10:313.
- Satoh, T. and M. Hosokawa. (2006) Chem.-Biol. Interactions, 162:195.
- Fleming, C. D. et al. (2007) Biochemistry 46:5603.
- Imai, T. (2006) Drug Metab. Pharmacokinet. 21:173.
Citations for Recombinant Human Carboxylesterase 2/CES2 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 3
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Discovery and molecular basis of subtype-selective cyclophilin inhibitors
Authors: AA Peterson, AM Rangwala, MK Thakur, PS Ward, C Hung, IR Outhwaite, AI Chan, DL Usanov, VK Mootha, MA Seeliger, DR Liu
Nature Chemical Biology, 2022-09-26;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Hydrolytic Fate of 3/15-Acetyldeoxynivalenol in Humans: Specific Deacetylation by the Small Intestine and Liver Revealed Using in Vitro and ex Vivo Approaches
Authors: el H Ajandouz, S Berdah, V Moutardier, T Bege, DJ Birnbaum, J Perrier, E Di Pasqual, M Maresca
Toxins (Basel), 2016-07-28;8(8):.
Species: Human
Sample Types: Complex Sample Type
Applications: Bioassay -
Mechanism of activation of PSI-7851 and its diastereoisomer PSI-7977.
Authors: Murakami E, Tolstykh T, Bao H, Niu C, Steuer HM, Bao D, Chang W, Espiritu C, Bansal S, Lam AM, Otto MJ, Sofia MJ, Furman PA
J. Biol. Chem., 2010-08-26;285(45):34337-47.
Species: Human
Sample Types: Small Molecule
Applications: Enzyme Assay
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