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 ...read more
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Recombinant Human DPPIV/CD26 (High Purity Dimer) 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 >8500 pmol/min/µg as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human DPPIV/CD26 protein Asn29-Pro766, with a C-terminal Asp-Ile and 6-His tag
>95%, 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
86 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
95-110 kDa, reducing condtions
Publications
Read Publications using 9168-SE in the following applications:
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.
Buffer
Supplied as a 0.2 μm filtered solution in MES and NaCl.
Purity
>95%, 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 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
Dilute rhDPPIV to 0.1 ng/μL in Assay Buffer.
Dilute Substrate to 200 μM in Assay Buffer.
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.
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 # A9891).
Per Well:
rhDPPIV: 0.005 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 DPPIV/CD26 (High Purity Dimer) Protein, CF
ADABP
ADCP-2
ADCP2DPP IV
Adenosine deaminase complexing protein 2TP103
CD26 antigen
CD26
CD26T-cell activation antigen CD26
dipeptidyl peptidase 4
Dipeptidyl peptidase IV
dipeptidylpeptidase 4
dipeptidyl-peptidase 4
dipeptidylpeptidase IV (CD26, adenosine deaminase complexing protein 2)
DPP4
DPPIV
EC 3.4.14.5
Background
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).
Klemann, C. et al. (2016) Clin. Exp. Immunol. Epub PMID 26919392.
Mortier, A. et al. (2016) J. Leukoc. Biol. Epub PMID 26744452.
Tanaka, T. et al. (1992) J. Immunol. 149:481.
Rohrborn, D. et al. (2014) FEBS Lett. 588:3870.
Waumans, Y. et al. (2015) Front. Immunol. 6:387.
Lamers, D. et al. (2011) Diabetes 60:1917.
Proost, P. et al. (1998) J. Biol. Chem. 273:7222.
Proost, P. et al. (2001) Blood 98:3554.
Ohtsuki, T. et al. (1998) FEBS Lett. 431:236.
Barreira da Silva, R. et al. (2015) Nat. Immunol. 16:850.
Guan, E. et al. (2002) J. Biol. Chem. 277:32348.
Callebaut, C. et al. (1993) Science 262:2045.
Raj, V.S. et al. (2013) Nature 495:251.
Broxmeyer, H.E. et al. (2012) Nat. Med. 18:1786.
Christopherson II, K.W. et al. (2004) Science 305:1000.
Kameoka, J. et al. (1993) Science 261:466.
Ohnuma, K. et al. (2007) J. Biol. Chem. 282:10117.
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