Recombinant Mouse Prolyl Oligopeptidase/PREP Protein, CF

Catalog # Availability Size / Price Qty
6339-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse Prolyl Oligopeptidase/PREP Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to convert the substrate benzyloxycarbonyl-Gly-Pro-7-amido-4-methylcoumarin (Z-GP-AMC) to Z-Gly-Pro and 7-amino-4-methylcoumarin (AMC). The specific activity is >4,500 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Prolyl Oligopeptidase/PREP protein
Leu2-Gln710, with an N-terminal Met and 6-His tag
Accession # Q9QUR6
Accession #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
82 kDa
SDS-PAGE
68-73 kDa, reducing conditions

Product Datasheets

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6339-SE

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.

6339-SE

Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and Glycerol.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 25 mM Tris, 250 mM NaCl, 2.5 mM DTT, pH 7.5
  • Recombinant Mouse Prolyl Oligopeptidase/PREP (rmPREP) (Catalog # 6339-SE)
  • Substrate: Z-GP-AMC (Bachem, Catalog # I-1145), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmPREP to 0.1 ng/μL in Assay Buffer.
  2. Dilute Substrate to 100 μM in Assay Buffer.
  3. Load into a plate 50 μL of 0.1 ng/μL rmPREP, and start the reaction by adding 50 μL of 100 μM Substrate.
  4. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 100 μM Substrate.
  5. Read plate at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:
  • rmPREP: 0.005 μg
  • Substrate: 50 μM
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: Prolyl Oligopeptidase/PREP

Prolyl Oligopeptidase, also known as prolyl endopeptidase and post-proline cleaving enzyme, is a serine peptidase displaying specificity for the cleavage of Pro-Xaa bonds of oligopeptide substrates (1, 2). The peptidase is known to hydrolyze a variety of biologically active peptides such as bradykinin, substance P, neurotensin, and vasopressin (3). Because of its action on neuropeptides, Prolyl Oligopeptidase is considered to be involved in processes such as learning, memory, and depression (4).

References
  1. Szeltner, Z and L. Polgár (2008) Current Protein Pept. Sci. 9:96.
  2. Yoshimoto, T. et al. (1977) Biochemistry. 16:2942.
  3. Wilk, S. (1983) Life Sci. 33:2149.
  4. Männisto, P.T. et al. (2007) Drug News Perspect. 20:293.
Entrez Gene IDs
5550 (Human); 19072 (Mouse); 83471 (Rat)
Alternate Names
dJ355L5.1 (prolyl endopeptidase); EC 3.4.21.26; MGC16060; PE; PEP; Post-proline cleaving enzyme; PREP; prolyl endopeptidase; Prolyl Oligopeptidase; rPop

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

FAQs

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