Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein, CF

R&D Systems | Catalog # 9168-SE

R&D Systems
Discontinued Product
9168-SE has been discontinued. An alternative/replacement product is available: 11244-SE. View all DPPIV/CD26 products.

Key Product Details

Source

NS0

Accession Number

CAA43118

Structure / Form

Noncovalently-linked homodimer

Applications

Enzyme Activity
View all DPPIV/CD26 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human DPPIV/CD26 protein
Asn29-Pro766, with a C-terminal Asp-Ile and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Asn29

Predicted Molecular Mass

86 kDa

SDS-PAGE

95-110 kDa, reducing condtions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Gly-Pro-7-amido-4-methylcoumarin (GP-AMC).
The specific activity is >8500 pmol/min/µg as measured under the described conditions.

Scientific Data Images for Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein, CF

Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein Enzyme Activity

Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein Enzyme Activity

Recombinant Human DPPIV/CD26 (Catalog # 9168-SE) is measured by its ability to cleave the fluorogenic peptide substrate, Gly-Pro-7-amido-4-methylcoumarin (GP-AMC). The activity (orange) is approximately 4-fold greater than the competitor's DPPIV/CD26 (green).

Formulation, Preparation, and Storage

9168-SE
Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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 opening.

Background: DPPIV/CD26

DPPIV/CD26 (EC 3.4.14.5) is an approximately 110 kDa serine exopeptidase that releases Xaa-Pro or Xaa-Ala dipeptides from the N-terminus of oligo- and polypeptides. It regulates immune and endocrine function through the cleavage of multiple chemokines, growth factors, and peptide hormones (1, 2). Mature human DPPIV consists of a 6 amino acid (aa) cytoplasmic tail, a 22 aa transmembrane segment, and a 738 aa extracellular domain (ECD) that contains the catalytic active site (Ser, Asp, and His charge relay system) (3). Within the ECD, human DPPIV/CD26 shares 84% amino acid sequence identity with mouse and rat DPPIV. DPPIV is expressed as a noncovalent homodimer on the surface of epithelial cells, endothelial cells, and activated lymphocytes, and it can be released by MMP mediated shedding (4). It cleaves a range of peptide hormones including Glucagon, Glucagon-like Peptides 1 and 2, GIP, GHRH, Procalcitonin, Neuropeptide Y, and Substance P (5). It is released from adipocytes and induces insulin resistance in adipocytes and skeletal muscle (6). DPPIV also cleaves many chemokines, resulting in reduced chemotactic activity of CXCL6, 9, 10, 11, 12, and CCL5 (7-10) but unchanged angiostatic activity of CXCL9 and CXCL10 (8). Cleavage can increase (CCL5), decrease (CXCL12), or have no effect (CCL4) on chemokine blockade of HIV-1 cellular infectivity (7, 9, 11). In addition, DPPIV cleavage of CCL4 broadens chemokine receptor usage to also include CCR2b (11). DPPIV serves as a cell entry coreceptor for HIV and coronavirus (12, 13). It cleaves human GM-CSF and IL-3 and reduces their ability to promote myeloid cell development (14). It also interferes with CXCL12 induced hematopoietic cell migration, homing, and engraftment (15). DPPIV interacts in cis with adenosine deaminase on T cells and in trans with caveolin-1 on antigen presenting cells (16, 17). It provides costimulatory proliferation and activation signals to both CD4+ and CD8+ T cells (17, 18).

References

  1. Klemann, C. et al. (2016) Clin. Exp. Immunol. Epub PMID 26919392.
  2. Mortier, A. et al. (2016) J. Leukoc. Biol. Epub PMID 26744452.
  3. Tanaka, T. et al. (1992) J. Immunol. 149:481.
  4. Rohrborn, D. et al. (2014) FEBS Lett. 588:3870.
  5. Waumans, Y. et al. (2015) Front. Immunol. 6:387.
  6. Lamers, D. et al. (2011) Diabetes 60:1917.
  7. Proost, P. et al. (1998) J. Biol. Chem. 273:7222.
  8. Proost, P. et al. (2001) Blood 98:3554.
  9. Ohtsuki, T. et al. (1998) FEBS Lett. 431:236.
  10. Barreira da Silva, R. et al. (2015) Nat. Immunol. 16:850.
  11. Guan, E. et al. (2002) J. Biol. Chem. 277:32348.
  12. Callebaut, C. et al. (1993) Science 262:2045.
  13. Raj, V.S. et al. (2013) Nature 495:251.
  14. Broxmeyer, H.E. et al. (2012) Nat. Med. 18:1786.
  15. Christopherson II, K.W. et al. (2004) Science 305:1000.
  16. Kameoka, J. et al. (1993) Science 261:466.
  17. Ohnuma, K. et al. (2007) J. Biol. Chem. 282:10117.
  18. Hatano, R. et al. (2013) Immunology 138:165.

Long Name

Dipeptidyl-peptidase IV

Alternate Names

CD26, DPP4

Entrez Gene IDs

1803 (Human); 13482 (Mouse); 397492 (Porcine); 102133935 (Cynomolgus Monkey)

Gene Symbol

DPP4

UniProt

CAA43118

Additional DPPIV/CD26 Products

Product Documents for Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein, CF

For research use only

Citations for Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein, CF

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Protocols

View specific protocols for Recombinant Human DPPIV/CD26 (High Purity Dimer) Protein, CF (9168-SE):

Materials
  • Assay Buffer: 25 mM Tris, pH 8.0
  • Recombinant Human DPPIV/CD26 (rhDPPIV) (Catalog # 9168-SE)
  • Substrate: H-Gly-Pro-AMC (Bachem, Catalog # I-1225), 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 rhDPPIV to 0.1 ng/μL in Assay Buffer.
  2. Dilute Substrate to 200 μM in Assay Buffer.
  3. Load into a black plate 50 μL of 0.1 ng/μL rhDPPIV, and start the reaction by adding 50 μL of 200 μM Substrate. As a Substrate Blank combine 50 μL of 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 # A9891).

Per Well:
  • rhDPPIV: 0.005 μg
  • Substrate: 100 μM






















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