Recombinant Mouse Carboxylesterase 1/CES1 Protein, CF
Recombinant Mouse Carboxylesterase 1/CES1 Protein, CF Summary
His19-Leu565, with a C-terminal 6-His tag
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.
|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 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
- Dilute rmCES1 to 12 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- 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.
- Incubate for 10 minutes at room temperature.
- Add 100 µL Reading Buffer to each well (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).
- rmCES1: 0.6 µg
- Substrate: 0.5 mM
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).
Redinbo, M.R. and Potter, P.M. (2005) Drug Discovery Today 10:313.
Satoh, T. and Hosokawa, M. (2006) Chem. Biol. Interactions 162:195.
Fleming, C.D. et al. (2007) Biochemistry 46:5603.
Ellinghaus P. et al. (1998) Biochim. Biophys. Acta. 1397:175.
Product Specific NoticesCoomassie is a registered trademark of Imperial Chemical Industries Ltd.
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