Dipeptidyl-peptidase II (DPPII), also known as quiescent cell proline dipeptidase (QPP) and DPP7, 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. They are all prolyl proteases that cleave proteins and peptides after proline residues. However, DPPII 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.
Applications
Suitable for use in ELISA, Neutralization and Western blot. Other applications have not been tested.
Recommended Dilutions
Neutralization-preincubated with the enzyme at different molar ratios at room temperature for 30 minutes. The enzyme was then assayed with a fluorogenic peptide substrate. 50% of the proteolytic activity was inhibited by the antibody at approximately 0.12ug/ml (IC50) under conditions in which the enzyme was present at 0.1ug/ml and the substrate concentration was 10uM. Considering the molecular masses of the enzyme (53kD) and the antibody (150kD), IC50 was achieved at approximately 1:2 molar ratio of the antibody to the enzyme. Western blot: 1-2ug/ml The detection limit is ~10 ng/lane under non-reducing conditions. Reducing conditions are not recommended. Chemiluminescent detection with WesternGloTM Chemiluminescent Detection Substrate will increase sensitivity by 5 to 50 fold. Optimal dilutions to be determined by the researcher.
Hybridoma
mouse myeloma with B cells obtained from a rat immunized with purified, NS0-derived, recombinant human DPPII.
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
NS0-derived recombinant human rhDPPII..
Form
Supplied as a lyophilized powder in PBS, 5% trehalose. Reconstitute with 200ul sterile PBS.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes human DPPII. Species sequence homology: chimpanzee 99%, mouse 81%, rat 80%, dog 79% and chicken 65%.