Recombinant Human DPP9 Protein, CF

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

Recombinant Human DPP9 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate, Gly-Pro-7-amido-4-methylcoumarin (GP-AMC). The specific activity is >750 pmol/min/μg, as measured under the described conditions.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human DPP9 protein
  Arg2-Leu892 with an N-terminal Met and 6-His tag
• Accession #: Q1ZZB8
• N-terminal Sequence: No result obtained
• Protein/Peptide Type: Recombinant Enzymes
• Gene: DPP9
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 101 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: 92 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 25 mM Tris, pH 8.0
  • Recombinant Human DPP9 (rhDPP9) (Catalog # 5419-SE)
  • Substrate: H-Gly-Pro-AMC (Bachem, Catalog # I-1225) Prepare a 10 mM stock in DMSO.
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhDPP9 to 1 ng/µL in Assay Buffer.
  2. Dilute substrate to 200 µM in Assay Buffer.
  3. Load 50 µL of 1 ng/µL rhDPP9 into a plate, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 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:
  • rhDPP9: 0.050 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 Human DPP9 Protein, CF

• dipeptidyl peptidase 9
• Dipeptidyl peptidase IV-related protein 2
• dipeptidyl peptidase IV-related protein-2
• Dipeptidyl peptidase IX
• Dipeptidyl peptidase-like protein 9
• dipeptidylpeptidase 9
• dipeptidyl-peptidase 9
• DKFZp762F117
• DP9
• DPLP9
• DPP IX
• DPP9
• DPRP2
• DPRP-2
• EC 3.4.14.5
• FLJ16073

Background

DPP9 is a member of the S9b family of serine peptidases (1, 2). It shares 19% amino acid identity with DPP4 and 58% amino acid identity with DPP8. It exhibits post‑proline dipeptidyl aminopeptidase activity, cleaving Xaa-Pro dipeptides from the N-terminus of oligo- and polypeptides (3). Unlike DPP4, DPP9 does not appear to be membrane bound and is localized exclusively in the cytoplasm (4). This family of proline-specific dipeptidyl peptidases has been implicated in a variety of diseases including type 2 diabetes, obesity and cancer, and has been a potential target for drug discovery (5, 6).

1. Olsen, C. and Wagtmann, N. (2002) Gene 299:185.
2. Qi, S.Y. et al. (2003) Biochem. J. 373:179.
3. Bjelke, J.R. et al. (2006) Biochem. J. 396:391.
4. Ajami, K. et al. (2004) Biochim. Biophys. Acta. 1679:18.
5. Rosenblum, J.S. and Kozarich, J.W. et al. (2003) Curr. Opin. Chem. Biol. 7:496.
6. Van der Veken, P. et al. (2007) Curr. Top. Med. Chem. 7:621.

Publications for DPP9 (5419-SE)(2)

Publications using 5419-SE

• JM Cho, EH Yang, W Quan, EH Nam, HG Cheon Discovery of a novel fibroblast activation protein (FAP) inhibitor, BR103354, with anti-diabetic and anti-steatotic effects Scientific Reports, 2020-12-04;10(1):21280. 2020-12-04 [PMID: 33277568] (Bioassay, N/A)
• DR Dunshee, TW Bainbridge, NM Kljavin, J Zavala-Sol, AC Schroeder, R Chan, R Corpuz, M Wong, W Zhou, G Deshmukh, J Ly, DP Sutherlin, JA Ernst, J Sonoda Fibroblast Activation Protein Cleaves and Inactivates Fibroblast Growth Factor 21 J. Biol. Chem, 2016-01-21;291(11):5986-96. 2016-01-21 [PMID: 26797127]

Bioinformatics

• Gene Symbol: DPP9
• Uniprot:
  • Q1ZZB8()