Recombinant Human DPPIV/CD26 Fc Chimera Protein, CF Summary
Accession # CAA43118.1
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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.
|Formulation||Supplied as a 0.2 μm filtered solution in Tris and NaCl.|
|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 Buffer: 25 mM Tris, pH 8.0
- Recombinant Human DPPIV/CD26 Fc Chimera (rhCD26/Fc) (Catalog # 11141-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 M5 by Molecular Devices) or equivalent
- Dilute rhCD26/Fc to 0.2 ng/μL in Assay Buffer.
- Dilute Substrate to 200 μM in Assay Buffer.
- Load into a black plate 50 μL of 0.2 ng/μL rhCD26/Fc, and start the reaction by adding 50 μL of 200 μM Substrate. For 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)|
- rhCD26/Fc: 0.010 μg
- Substrate: 100 μM
Recombinant Human DPPIV/CD26 Fc Chimera (Catalog # 11141-SE) is measured by its ability to cleave the fluorogenic peptide substrate, Gly-Pro-7-amido-4-methylcoumarin (GP-AMC).
2 μg/lane of Recombinant Human DPPIV/CD26 Fc Chimera Protein (Catalog # 11141-SE) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 116-131 kDa and 230-260 kDa, respectively.
Dipeptidyl peptidase 4 (DPPIV, also known as CD26) is an approximately 110 kDa serine exopeptidase that releases Xaa-Pro or Xaa-Ala dipeptides from the N-terminus of oligo- and polypeptides. 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 (1). 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 (2). It regulates immune and endocrine function through the cleavage of multiple chemokines, growth factors, and peptide hormones (3,4). 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 altered chemotactic activity (7-10) or impacting chemokine blockade of HIV-1 cellular infectivity depending on the chemokine target (7,9,11). It cleaves human GM-CSF and IL-3 and reduces their ability to promote myeloid cell development (12). In addition to enzymatic cleavage functions, DPPIV interacts with adenosine deaminase on T cells and with caveolin-1 on antigen presenting cells (13), provides costimulatory proliferation and activation signals to both CD4+ and CD8+ T cells (13,14), and serves as a cell entry coreceptor for HIV and coronavirus MERS-CoV-2 (15,16). DPPIV inhibitors are approved for therapeutic treatment in type 2 diabetes (17) and are being explored for treatment of several additional conditions including autoimmune diseases (18).
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