Recombinant Mouse DPP7 Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse DPP7 Protein, CF Summary

• Details of Functionality: 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 #: NP_114031
• N-terminal Sequence: Asp34
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Dpp7
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 54 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 65 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 200 μg/mL in 25 mM Tris and 150 mM NaCl, pH 7.5.
• Assay Procedure:
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse DPP7 Protein, CF

• 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

Background

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.

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.

Bioinformatics

• Gene Symbol: Dpp7
• Uniprot:
  • NP_114031()