Recombinant Human Prolyl Oligopeptidase/PREP Protein, CF Summary
Leu2-Pro710, with an N-terminal Met and 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 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.
- 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
- Dilute rhPREP to 0.1 µg/mL in Assay Buffer.
- Dilute Substrate to 100 µM in Assay Buffer.
- Incubate at room temperature for 5 minutes.
- 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.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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).
- rhPREP: 0.005 µg
- Substrate: 50 µM
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).
- Tarrago, T. et al. (2005) J. Pept. Sci. 11:283.
- Yoshimoto, T. et al. (1977) Biochemistry. 16:2942.
- Wilk, S. (1983) Life Sci. 33:2149.
- Maes, M. et al. (1994) Biol. Psychiatry. 35:545.
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.
Citations: Showing 1 - 3
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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;10(1):21280.
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;13(1):47.
Sample Types: Peptide
Applications: Enzyme Assay
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;0(0):.
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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