Recombinant Human Prolyl Oligopeptidase/PREP Protein, CF

Catalog # Availability Size / Price Qty
4308-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (5)
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Recombinant Human Prolyl Oligopeptidase/PREP Protein, CF Summary

Product Specifications

Purity
>95%, 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 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 >3,500 pmol/min/µg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Prolyl Oligopeptidase/PREP protein
Leu2-Pro710, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
81 kDa
SDS-PAGE
73 kDa, reducing conditions

Product Datasheets

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4308-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.

4308-SE

Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and Glycerol.
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, -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 Human Prolyl Oligopeptidase/PREP (rhPREP) (Catalog # 4308-SE)
  • Substrate: Z-Gly-Pro-AMC (Bachem, Catalog # I-1145)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhPREP to 0.1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 100 µM in Assay Buffer.
  3. Incubate at room temperature for 5 minutes.
  4. Load 50 µL of 0.1 µg/mL of rhPREP into a plate, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  5. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), 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 (AMC) (Sigma, Catalog # A-9891).

Per Well:
  • rhPREP: 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 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. Tarrago, T. et al. (2005) J. Pept. Sci. 11:283.
  2. Yoshimoto, T. et al. (1977) Biochemistry. 16:2942.
  3. Wilk, S. (1983) Life Sci. 33:2149.
  4. Maes, M. et al. (1994) Biol. Psychiatry. 35:545.
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

Citations for Recombinant Human Prolyl Oligopeptidase/PREP 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.

5 Citations: Showing 1 - 5
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  1. Isolation and characterization of natural inhibitors of post-proline specific peptidases from the leaves of Cotinus coggygria Scop
    Authors: Ivanov, I;Vasileva, A;Tasheva, D;Dimitrova, M;
    Journal of ethnopharmacology
    Species: Human
    Sample Types: Small Molecule
    Applications: Bioassay
  2. Discovery of a novel fibroblast activation protein (FAP) inhibitor, BR103354, with anti-diabetic and anti-steatotic effects
    Authors: JM Cho, EH Yang, W Quan, EH Nam, HG Cheon
    Scientific Reports, 2020-12-04;10(1):21280.
    Species: N/A
    Sample Types:
    Applications: Bioassay
  3. Alzheimer-associated cerebrospinal fluid fragments of neurogranin are generated by Calpain-1 and prolyl endopeptidase
    Authors: B Becker, FH Nazir, G Brinkmalm, E Camporesi, H Kvartsberg, E Portelius, M Boström, M Kalm, K Höglund, M Olsson, H Zetterberg, K Blennow
    Mol Neurodegener, 2018-08-29;13(1):47.
    Species: Human
    Sample Types: Peptide
    Applications: Enzyme Assay
  4. Pericyte-targeting prodrug overcomes tumor resistance to vascular disrupting agents
    Authors: M Chen, X Lei, C Shi, M Huang, X Li, B Wu, Z Li, W Han, B Du, J Hu, Q Nie, W Mai, N Ma, N Xu, X Zhang, C Fan, A Hong, M Xia, L Luo, A Ma, H Li, Q Yu, H Chen, D Zhang, W Ye
    J. Clin. Invest., 2017-08-28;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  5. Fibroblast Activation Protein Cleaves and Inactivates Fibroblast Growth Factor 21
    Authors: DR Dunshee, TW Bainbridge, NM Kljavin, J Zavala-Sol, AC Schroeder, R Chan, R Corpuz, M Wong, W Zhou, G Deshmukh, J Ly, DP Sutherlin, JA Ernst, J Sonoda
    J. Biol. Chem, 2016-01-21;291(11):5986-96.

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