Recombinant Mouse Prolyl Oligopeptidase/PREP Protein, CF

Catalog #: 6339-SE Datasheet / COA / SDS

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6339-SE-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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 #

Q9QUR6

N-terminal Sequence
Analysis

Met

Predicted Molecular Mass

82 kDa

SDS-PAGE

68-73 kDa, reducing conditions

Product Datasheets

6339-SE

Product Datasheet

COA

SDS

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

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 V_max* (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

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

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.

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