The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 24466060
1/1000 - 1/5000. Detects a band of approximately 110-120 kDa (predicted molecular weight: 88 kDa). 1/1000 when using colorimetric substrates such as BCIP/NBT - 1/5000 for chemiluminescent substrates. Bands run high because of post-translational modifications and reduction. EDTA/EGTA treatment of tissues or lysates is required to see latent zymogen. Dilution optimised using Chromogenic detection.
Use at an assay dependent concentration.
Cell surface glycoprotein receptor involved in the costimulatory signal essential for T-cell receptor (TCR)-mediated T-cell activation. Acts as a positive regulator of T-cell coactivation, by binding at least ADA, CAV1, IGF2R, and PTPRC. Its binding to CAV1 and CARD11 induces T-cell proliferation and NF-kappa-B activation in a T-cell receptor/CD3-dependent manner. Its interaction with ADA also regulates lymphocyte-epithelial cell adhesion. In association with FAP is involved in the pericellular proteolysis of the extracellular matrix (ECM), the migration and invasion of endothelial cells into the ECM. May be involved in the promotion of lymphatic endothelial cells adhesion, migration and tube formation. When overexpressed, enhanced cell proliferation, a process inhibited by GPC3. Acts also as a serine exopeptidase with a dipeptidyl peptidase activity that regulates various physiological processes by cleaving peptides in the circulation, including many chemokines, mitogenic growth factors, neuropeptides and peptide hormones. Removes N-terminal dipeptides sequentially from polypeptides having unsubstituted N-termini provided that the penultimate residue is proline.
Expressed specifically in lymphatic vessels but not in blood vessels in the skin, small intestine, esophagus, ovary, breast and prostate glands. Not detected in lymphatic vessels in the lung, kidney, uterus, liver and stomach (at protein level). Expressed in the poorly differentiated crypt cells of the small intestine as well as in the mature villous cells. Expressed at very low levels in the colon.
Belongs to the peptidase S9B family. DPPIV subfamily.
The extracellular cysteine-rich region is necessary for association with collagen, dimer formation and optimal dipeptidyl peptidase activity.
The soluble form (Dipeptidyl peptidase 4 soluble form also named SDPP) derives from the membrane form (Dipeptidyl peptidase 4 membrane form also named MDPP) by proteolytic processing. N- and O-Glycosylated. Phosphorylated. Mannose 6-phosphate residues in the carbohydrate moiety are necessary for interaction with IGF2R in activated T-cells. Mannose 6-phosphorylation is induced during T-cell activation.
Cell membrane. Apical cell membrane. Cell projection > invadopodium membrane. Cell projection > lamellipodium membrane. Cell junction. Membrane raft. Translocated to the apical membrane through the concerted action of N- and O-Glycans and its association with lipid microdomains containing cholesterol and sphingolipids. Redistributed to membrane rafts in T-cell in a interleukin-12-dependent activation. Its interaction with CAV1 is necessary for its translocation to membrane rafts. Colocalized with PTPRC in membrane rafts. Colocalized with FAP in invadopodia and lamellipodia of migratory activated endothelial cells in collagenous matrix. Colocalized with FAP on endothelial cells of capillary-like microvessels but not large vessels within invasive breast ductal carcinoma. Colocalized with ADA at the cell junction in lymphocyte-epithelial cell adhesion. Colocalized with IGF2R in internalized cytoplasmic vesicles adjacent to the cell surface and Secreted. Detected in the serum and the seminal fluid.
Postranslational modifications and reduction lead to an apparent mass of 110-120 kDa on SDS-PAGE gels.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD26 antibody (ab28340)This image is a courtesy of Veronique Dubois
ab28340 staining CD26 - Spacer region in rat epididymis tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent paraformaldehyde fixation before heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 and then blocking with 3% hydrogen peroxide in TBS for 10 minutes at 25°C was performed in order to block endogenous peroxidase activity. The primary antibody was used diluted in 1/400 and it was incubated with sample for 16 hours at 4°C in a dilution buffer containing TBS, 0.3% Triton X-100, 0.1 % BSA. A Biotin conjugated goat polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CD26 antibody (ab28340)Image from Gieseck RL 3rd et al., PLoS One. 2014;9(1):e86372. Fig 1c.; doi: 10.1371/journal.pone.0086372.
Immunocytochemistry/Immunofluorscence analysis of IPSC cells labelling CD26 with ab28340 at a dilution of 1/100. Cells were fixed for 30 minutes at 4°C in 4% paraformaldehyde and washed 3 time with DPBS. Cells were blocked for 1 hours with DPBS containing 1% donkey serum and 1% Triton X-100. Cells were incubated for 1 hour at room temperature with ab28340 and ab32572 (anti-beta Catenin, 1/100) in blocking buffer. Cells were washed three times with PBS for 30 minutes each. Cells were incubated for 1 hour at room temperature with appropriate secondary antibodies diluted in the blocking solution. Cells were then washed three times with PBS for 30 minutes each and then imaged using an LSM700 laser scanning confocal microscope.