Recombinant Mouse DPP7 Protein, CF Summary
Product Specifications
Asp34-Arg506, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
|
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
- Dilute rmDPPII to 0.04 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- 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.
- Read 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).
- rmDPPII: 0.002 µg
- Substrate: 100 µM
Reconstitution Calculator
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.
- Araki, H. et al. (2001) J. Biochem. 129:279.
- Maes, M.-B. et al. (2005) Biochem. J. 386:315.
- Rosenblum, J.S. and J.W. Kozarich (2003) Curr. Opin. Chem. Biol. 7:496.
- Lankas, G.R. et al. (2005) Diabetes 54:2988.
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