Recombinant Mouse Carboxyl Ester Lipase/CEL Protein, CF

Catalog # Availability Size / Price Qty
5658-CE-010
Product Details
Citations (1)
FAQs
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Recombinant Mouse Carboxyl Ester Lipase/CEL Protein, CF Summary

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave p-Nitrophenyl butyrate (PNPB). The specific activity is >4,000 pmol/min/μg, as measured under the described conditions. 
Source
Mouse myeloma cell line, NS0-derived mouse Carboxyl Ester Lipase/CEL protein
Ala21-Leu534, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Ala21
Structure / Form
Monomer
Predicted Molecular Mass
58 kDa
SDS-PAGE
60 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

5658-CE

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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. Dilute rmCEL to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. 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.
  4. Incubate 10 minutes at room temperature.
  5. Quickly add 100 µL of Reading Buffer to all wells (this will not stop the reaction).
  6. Read immediately at a wavelength of 410 nm (bottom read) in endpoint mode.
  7. 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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References
  1. Wang, C.S. and J.A. Hartsuck, J.A. (1993) Biochim. Biophys. Acta. 1166:1.
  2. Lombardo, D. (2001) Biochim. Biophys. Acta. 1533:1.
  3. Hui, D.Y. and Howles, P.N. (2002) J. Lipid Res. 43:2017.
  4. Brodt-Eppley, J. et al. (1995) Biochim. Biophys. Acta. 1272:69.
Entrez Gene IDs
1056 (Human); 12613 (Mouse); 24254 (Rat)
Alternate Names
BAL; bile salt-activated lipase; Bile salt-stimulated lipase; BSDL; BSSL; BSSLbile salt-dependent lipase, oncofetal isoform; bucelipase; carboxyl ester hydrolase; carboxyl ester lipase (bile salt-stimulated lipase); Carboxyl Ester Lipase; CEase; CEL; CELL; Cholesterol esterase; EC 3.1.1; EC 3.1.1.13; EC 3.1.1.3; FAP; FAPP; fetoacinar pancreatic protein; LIPA; lysophospholipase, pancreatic; MODY8bile-salt-activated lipase; Pancreatic lysophospholipase; Sterol esterase

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

  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.
    Species: Human
    Sample Types:
    Applications: Western Blot

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