Recombinant Mouse Carboxylesterase 31-like/EG13909, CF Summary
Met1-Glu558, with a C-terminal 10-His tag
Accession # Q8VCU1
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, pH 7.5
- Absorbance Buffer: 50 mM Tris, pH 8.0
- Recombinant Mouse Carboxylesterase 31 like/EG13909 (rmEG13909) (Catalog # 6059-CE)
- Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rmEG13909 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 2.0 mM in Assay Buffer.
- Load 50 µL of 2 ng/µL rmEG13909 into plate and start the reaction by adding 50 µL of 2.0 mM Substrate. As a Substrate Blank combine 50 µL Assay Buffer with 50 µL Substrate.
- Cover and incubate at room temperature for 60 minutes in the dark.
- After incubation add 100 µL Absorbance Buffer and read plate immediately.
- Read (top read) absorbance in endpoint mode at 410 nm.
- 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 p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).
- rmEG13909: 0.1 µg
- Substrate: 0.5 mM
Background: Carboxylesterase 31-like/EG13909
EG13909, also known as Es-31, is a carboxylesterase-like enzyme identified in mouse liver (1). It belongs to the type B carboxylesterase/lipase superfamily that hydrolyzes the carboxylic ester bonds. By sequence analysis it is most closely related to mouse CES-3 and is likely to be a microsomal enzyme. It has been suggested that EG13909 is expressed predominantly in male livers and might be regulated by growth hormone (1).
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