Recombinant Mouse Carboxyl Ester Lipase/CEL Protein, CF Summary
Ala21-Leu534, with a C-terminal 6-His tag
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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 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 Buffer: 25 mM Tris, pH 7.0
- Reading Buffer: 25 mM Tris, pH 8.0
- Recombinant Mouse Carboxyl Ester Lipase/CEL (rmCEL) (Catalog # 5658-CE)
- Substrate: p-Nitrophenyl butyrate (PNPB) (Sigma, Catalog # N9876), 100 mM stock in acetone
- 96-well Clear Plate (Catalog # DY990)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rmCEL to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load into the wells of a clear microplate 50 μL of 0.2 ng/µL rmCEL, and start the reaction by adding 50 µL of Substrate. Include a Blank Control containing 50 µL of Assay Buffer and 50 µL of Substrate without any rmCEL.
- Incubate 10 minutes at room temperature.
- Quickly add 100 µL of Reading Buffer to all wells (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).Per Well:
- rmCEL: 0.010 µg
- Substrate: 0.5 mM
Background: Carboxyl Ester Lipase/CEL
Carboxyl ester lipase, also known as bile salt-stimulated lipase, belongs to the Type-B carboxyl esterase/lipase family of enzymes. The enzyme possesses a Ser‑His‑Asp catalytic triad, and is capable of hydrolyzing both ester and amide bonds. CEL is a non-specific lipase which hydrolyzes a variety of substrates, including cholesteryl esters, acylglycerols, and ceramide (1). CEL is highly expressed by pancreatic acinar cells and secreted into the gastrointestinal tract, where it contributes to the digestion of dietary lipids (2). It is expressed at much lower levels by the liver, macrophages, and endothelial cells (3). CEL is present in the circulation, where it may play a role in low density lipoprotein metabolism and atherogenesis (4).
- Wang, C.S. and J.A. Hartsuck, J.A. (1993) Biochim. Biophys. Acta. 1166:1.
- Lombardo, D. (2001) Biochim. Biophys. Acta. 1533:1.
- Hui, D.Y. and Howles, P.N. (2002) J. Lipid Res. 43:2017.
- Brodt-Eppley, J. et al. (1995) Biochim. Biophys. Acta. 1272:69.
Citation for Recombinant Mouse Carboxyl Ester Lipase/CEL 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.
1 Citation: Showing 1 - 1
A recombined allele of the lipase gene CEL and its pseudogene CELP confers susceptibility to chronic pancreatitis.
Authors: Fjeld K, Weiss F, Lasher D, Rosendahl J, Chen J, Johansson B, Kirsten H, Ruffert C, Masson E, Steine S, Bugert P, Cnop M, Grutzmann R, Mayerle J, Mossner J, Ringdal M, Schulz H, Sendler M, Simon P, Sztromwasser P, Torsvik J, Scholz M, Tjora E, Ferec C, Witt H, Lerch M, Njolstad P, Johansson S, Molven A
Nat Genet, 2015;47(5):518-22.
Applications: Western Blot
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