Recombinant Human DPPIV/CD26 Protein, CF Summary
Product Specifications
Asn29-Pro766, with a C-terminal Asp-Ile and 6-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.
1180-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.
|
Assay Procedure
- Assay Buffer: 25 mM Tris, pH 8.0
- Recombinant Human DPPIV/CD26 (rhDPPIV) (Catalog # 1180-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 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load into a black plate 50 µL of 0.2 ng/µL rhDPPIV and start the reaction by adding 50 µL of 20 µM Substrate. As a Substrate Blank combine 50 µL of Assay Buffer and 50 µL of 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)
Per Well:- rhDPPIV: 0.010 µg
- Substrate: 10 µM
Scientific Data
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Recombinant Human DPPIV/CD26 (Catalog # 1180-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 2-fold greater than the competitor's DPPIV/CD26 (green).
Reconstitution Calculator
Background: DPPIV/CD26
DPPIV/CD26 (EC 3.4.14.5) is a serine exopeptidase that releases Xaa-Pro dipeptides from the N-terminus of oligo- and polypeptides (1, 2). It is a type II membrane protein consisting of a short cytoplasmic tail, a transmembrane domain, and a long extracellular domain (3‑5). The extracellular domain contains glycosylation sites, a cysteine-rich region and the catalytic active site (Ser, Asp and His charge relay system). The amino acid sequence of the mouse DPPIV/CD26 extracellular domain is 84% and 91% identical to the human and rat counterparts, respectively. In the native state, DPPIV/CD26 is present as a noncovalently linked homodimer on the cell surface of a variety of cell types. The soluble form is also detectable in human serum and other body fluids, the levels of which may have clinical significance in patients with cancer, liver and kidney diseases, and depression.
DPPIV/CD26 plays an important role in many biological and pathological processes. It functions as T cell-activating molecule (THAM). It serves as a cofactor for entry of HIV in CD4+ cells (6). It binds adenosine deaminase, the deficiency of which causes severe combined immunodeficiency disease in humans (7). It cleaves chemokines such as stromal-cell-derived factor 1 alpha and macrophage-derived chemokine (8, 9). It degrades peptide hormones such as glucagon (10). It truncates procalcitonin, a marker for systemic bacterial infections with elevated levels detected in patients with thermal injury, sepsis and severe infection, and in children with bacterial meningitis (11).
- Misumi and Ikehara (1998) in Handbook of Proteolytic Enzymes. Barrett, et al. (eds) San Diego: Academic Press, p. 378.
- Ikehara, et al. (1994) Methods Enzymol. 244:215.
- Marguet, et al. (1992) J. Biol. Chem. 267:2200.
- Bernard, et al. (1994) Biochemistry 33:15204.
- Viver, et al. (1991) J. Immunol. 147:447.
- Callebaut, et al. (1993) Science 262:2045.
- Kameoka, et al. (1993) Science 261:466.
- Ohtsuki, et al. (1998) FEBS Lett. 431:236.
- Proost, et al. (1999) J. Biol. Chem. 274:3988.
- Hinke, et al. (2000) J. Biol. Chem. 275:3827.
- Wrenger, et al. (2000) FEBS Lett. 466:155.
Citations for Recombinant Human DPPIV/CD26 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 4
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Elevated hepatic DPP4 activity promotes insulin resistance and non-alcoholic fatty liver disease
Authors: C Baumeier, L Schlüter, S Saussentha, T Laeger, M Rödiger, SA Alaze, L Fritsche, HU Häring, N Stefan, A Fritsche, RW Schwenk, A Schürmann
Mol Metab, 2017-08-04;6(10):1254-1263.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Fibroblast Activation Protein Cleaves and Inactivates Fibroblast Growth Factor 21
Authors: 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
J. Biol. Chem, 2016-01-21;291(11):5986-96.
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Human Opiorphin, a natural antinociceptive modulator of opioid-dependent pathways.
Authors: Wisner A, Dufour E, Messaoudi M, Nejdi A, Marcel A, Ungeheuer MN, Rougeot C
Proc. Natl. Acad. Sci. U.S.A., 2006-11-13;103(47):17979-84.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
E3024, 3-but-2-ynyl-5-methyl-2-piperazin-1-yl-3,5-dihydro-4H-imidazo[4,5-d]pyridazin-4-one tosylate, is a novel, selective and competitive dipeptidyl peptidase-IV inhibitor.
Authors: Yasuda N, Nagakura T, Inoue T, Yamazaki K, Katsutani N, Takenaka O, Clark R, Matsuura F, Emori E, Yoshikawa S, Kira K, Ikuta H, Okada T, Saeki T, Asano O, Tanaka I
Eur. J. Pharmacol., 2006-08-16;548(1):181-7.
Species: Human
Sample Types: Pharmaceutical
Applications: Enzyme Assay
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