Recombinant Mouse Prolyl Oligopeptidase/PREP Protein, CF Summary
Leu2-Gln710, with an N-terminal Met and 6-His tag
Accession # Q9QUR6
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 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.
- 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
- Dilute rmPREP to 0.1 ng/μL in Assay Buffer.
- Dilute Substrate to 100 μM in Assay Buffer.
- Load into a plate 50 μL of 0.1 ng/μL rmPREP, and start the reaction by adding 50 μL of 100 μM Substrate.
- For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 100 μM Substrate.
- Read plate at excitation and emission wavelengths of 380 nm and 460 nm, 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 (Sigma, Catalog # A-9891).
- rmPREP: 0.005 μg
- Substrate: 50 μM
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).
- Szeltner, Z and L. Polgár (2008) Current Protein Pept. Sci. 9:96.
- Yoshimoto, T. et al. (1977) Biochemistry. 16:2942.
- Wilk, S. (1983) Life Sci. 33:2149.
- Männisto, P.T. et al. (2007) Drug News Perspect. 20:293.
Product Specific NoticesCoomassie is a registered trademark of Imperial Chemical Industries Ltd.
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