Recombinant Mouse DPP7 Protein, CF

Catalog # Availability Size / Price Qty
3436-SE-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Mouse DPP7 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 cleave the fluorogenic peptide substrate, Lys-Pro-AMC (KP-AMC). The specific activity is >20,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse DPPII/QPP/DPP7 protein
Asp34-Arg506, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Asp34
Predicted Molecular Mass
54 kDa
SDS-PAGE
65 kDa, reducing conditions

Product Datasheets

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.

3436-SE

Formulation Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Reconstitution Reconstitute at 200 μg/mL in 25 mM Tris and 150 mM NaCl, pH 7.5.
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 reconstitution.

Assay Procedure

Materials
  • Assay Buffer: 25 mM MES, pH 6.0
  • Recombinant Mouse DPPII/QPP/DPP7 (rmDPPII) (Catalog # 3436-SE)
  • Substrate: Lys-Pro-AMC (Bachem, Catalog # 1745), 10 mM stock solution in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmDPPII to 0.04 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load into plate 50 µL of 0.04 ng/µL rmDPPII, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
  5. 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:
  • rmDPPII: 0.002 µg
  • Substrate: 100 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: DPPII/QPP/DPP7

Dipeptidyl-peptidase II (DPPII) is identical to quiescent cell proline dipeptidase (QPP) and dipeptidylpeptidase 7 (DPP7) (1, 2). It shares some substrate and cleavage specificity with DPPIV/CD26, DPP8, DPP9 and seprase/FAP (fibroblast activation protein), members of the S09 family of serine proteases. As prolyl proteases that cleave proteins and peptides after proline residues, these enzymes have a high potential for drug discovery (3, 4). However, DPP7 is not a member of the S09 family, but a member of the S28 family that also includes lysosomal Pro-X carboxypeptidase/prolylcarboxypeptidase/PRCP and thymus-specific serine peptidase/PRSS16 (2). The mouse DPP7 precursor consists of a  signal peptide (amino acids (aa) 1-33) and a mature chain (aa 34-506). The purified recombinant mouse DPP7 is active against Lys-Pro-AMC and Lys-Ala-AMC. Its activity against Lys-Pro-AMC is approximately 10-fold of that against Lys-Ala-AMC under otherwise identical conditions.

References
  1. Araki, H. et al. (2001) J. Biochem. 129:279.
  2. Maes, M.-B. et al. (2005) Biochem. J. 386:315.
  3. Rosenblum, J.S. and J.W. Kozarich (2003) Curr. Opin. Chem. Biol. 7:496.
  4. Lankas, G.R. et al. (2005) Diabetes 54:2988.
Long Name
Dipeptidyl-peptidase II
Entrez Gene IDs
29952 (Human); 83768 (Mouse); 83799 (Rat)
Alternate Names
carboxytripeptidase; Dipeptidyl aminopeptidase II; dipeptidyl arylamidase II; dipeptidyl peptidase 2; Dipeptidyl peptidase 7; Dipeptidyl peptidase II; dipeptidylpeptidase 7; dipeptidyl-peptidase 7; dipeptidyl-peptidase II; DPP II; DPP2; DPP7; DPPII; EC 3.4.14.2; QPP; Quiescent cell proline dipeptidase

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