polyclonal goat igg af1180  (R&D Systems)

Journal: JID Innovations

Article Title: Human keratinocytes exhibit limited potential for SARS-CoV-2 infection despite ACE2 and mature cathepsin L expression

doi: 10.1016/j.xjidi.2025.100447

Figure Lengend Snippet: Differential ACE2, TMPRSS2, NRP1, CTSL, CD147, AXL, and DPP4 protein expression. Whole-cell lysates of the indicated cells were analyzed by western blot using the following antibodies: ACE2 (mAb clone AC384), TMPRSS2 (mAb clone S20014A), NRP1 (mAb clone 14H4), CTSL (mAb clone 33/1), CD147 (mAb clone HIM6), AXL (polyclonal goat IgG, AF154), and DPP4 (polyclonal goat IgG, AF1180). ( a ) Representative immunoblots from 2 to 4 independent experiments. ( b ) Densitometric quantification normalized to actin, expressed as mean ± SEM from 2 to 4 independent experiments. CTSL, cathepsin L.

Article Snippet: Primary antibodies included anti-ACE2, clone AC384 (AdipoGen Life Science/Coger, Paris, France); anti-TMPRSS2, clone S20014A; anti-NRP1, clone 14H4; anti-CD147, clone HIM6; anti–keratin 10, rabbit polyclonal Poly19054 (BioLegend); anti-CTSL, clone 33/1 (eBioscience, Thermo Fisher Scientific); anti-AXL, polyclonal goat IgG AF154; and anti-DPP4, polyclonal goat IgG AF1180 (R&D Systems, Bio-Techne SAS, Noyal Châtillon, France).

Techniques: Expressing, Western Blot

Journal: iScience

Article Title: Pancreas-derived DPP4 is not essential for glucose homeostasis under metabolic stress.

doi: 10.1016/j.isci.2023.106748

Figure Lengend Snippet: Figure 2. Islets from HFD-fed Dpp4b-cell/ mice have significantly reduced islet DPP4 activity and expression (A) mRNA abundance of Dpp4 in islets isolated from 52-week-old wild-type C57BL/6J mice fed chow or a high-fat diet. (B) DPP4 activity normalized to total protein in isolated islets from young female mice fed chow or high-fat diet (5 weeks). (C and D) Active GLP-1 and insulin in perfusate from islets of WT male and female mice fed chow or HFHC (12 weeks) diets exposed to 16.7 mM and 10 mM glucose in dynamic perifusion. (E) Insulin content measured in the pancreas of 12-week-old Dpp4+/+ or Dpp4/ littermate controls fed chow or HFD (4 weeks). (F) DPP4 activity normalized to total protein measured in whole extracts of tissues (liver, spleen, gut, heart, pancreas, islets) control (WT, MIP-Cre, Dpp4fl/fl) and Dpp4b-cell/ male mice. (G and H) Plasma DPP4 activity (G) and concentration (H) in plasma of HFD-fed control and Dpp4b-cell/ male mice.

Article Snippet: Sections were incubated with polyclonal goat anti-mouse DPP4 (1/40, R&D Systems, AF954), recombinant rabbit monoclonal anti-insulin (1/250; Abcam ab181547, clone number EPR17359), and recombinant anti-glucagon (1/500; Abcam ab92517, clone number EP3070) primary anti- bodies overnight at 4 C. Sections were incubated with Alexa Fluor488 donkey anti-goat (1/500; Thermo Fisher Scientific A32814) and Alexa Fluor Plus 647 donkey anti-rabbit (1/500; Thermo Fisher Scientific A32795) secondary antibodies for 45 minutes and with DAPI for 5 minutes protected from light at room temperature.

Techniques: Activity Assay, Expressing, Isolation, Control, Clinical Proteomics, Concentration Assay

Journal: iScience

Article Title: Pancreas-derived DPP4 is not essential for glucose homeostasis under metabolic stress.

doi: 10.1016/j.isci.2023.106748

Figure Lengend Snippet: Figure 3. Elimination of b cell-derived DPP4 does not improve glucose tolerance and insulin tolerance or prevent incretin degradation in male HFD-fed mice (A–D) Oral glucose tolerance (A) and plasma active GIP (B), active GLP-1 (C), and insulin (D) in male HFD-fed Dpp4b-cell/ and MIP-Cre control mice in response to oral glucose gavage G sitagliptin. (E–G) Intraperitoneal (i.p.) glucose tolerance (E) and plasma active GIP (F), and insulin (G) secreted in response to i.p. glucose injection. Sitagliptin was given by oral gavage 30 min prior to glucose tolerance tests (2 g/kg body weight). (H) Glucose levels and AUC during an insulin (0.6 IU/kg) tolerance test in male, HFD-fed mice. (I) Insulin tolerance test glycemia as a percentage of fasted glucose and AUC. (J) Insulin secretion measured in perifusion of islets from 65-week-old HFD-fed Dpp4b-cell/ (n = 4) and MIP-Cre control (n = 4) mice with arginine (1 mM), GLP-1 (0.3 mm) and GIP (100 nM). Graphs were analyzed by one-way ANOVA (A, E AUCs), unpaired t test (H, I AUCs), or mixed-effects analysis with Tukey’s multiple comparisons. All data are represented as the mean G SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Sections were incubated with polyclonal goat anti-mouse DPP4 (1/40, R&D Systems, AF954), recombinant rabbit monoclonal anti-insulin (1/250; Abcam ab181547, clone number EPR17359), and recombinant anti-glucagon (1/500; Abcam ab92517, clone number EP3070) primary anti- bodies overnight at 4 C. Sections were incubated with Alexa Fluor488 donkey anti-goat (1/500; Thermo Fisher Scientific A32814) and Alexa Fluor Plus 647 donkey anti-rabbit (1/500; Thermo Fisher Scientific A32795) secondary antibodies for 45 minutes and with DAPI for 5 minutes protected from light at room temperature.

Techniques: Derivative Assay, Clinical Proteomics, Control, Injection

Journal: iScience

Article Title: Pancreas-derived DPP4 is not essential for glucose homeostasis under metabolic stress.

doi: 10.1016/j.isci.2023.106748

Figure Lengend Snippet: Figure 4. Elimination of b cell-derived DPP4 does not improve glucose tolerance, incretin levels, or GSIS in HFD-fed female mice (A–C) Oral glucose tolerance (A), active GIP (B), and insulin secreted (C) in female HFD-fed Dpp4b-cell/ and MIP-Cre control mice in response to oral glucose gavage G sitagliptin. (D–F) Intraperitoneal (i.p.) glucose tolerance (D), active GIP (E), and insulin secreted (F) in female HFD-fed Dpp4b-cell/ and MIP-Cre control mice in response to i.p. glucose injection. Sitagliptin was given by oral gavage 30 min prior to glucose tolerance tests (2 g/kg body weight for oGTT and at 1.2 g/kg for ipGTT) (n = 7–10 per group). (G) Glucose levels and AUC during an insulin (0.6 IU/kg) tolerance test in female HFD-fed Dpp4b-cell/ and MIP-Cre control mice. (H) Insulin tolerance test glycemia as a percentage of fasted glucose and AUC. (I) GSIS measured during perifusion of islets isolated from 65-week-old HFD-fed Dpp4b-cell/ (n = 4) and MIP-Cre control (n = 4) mice with arginine (1 mM), GLP-1 (0.3 mm) and GIP (100 nM). AUC graphs represent area under the curve, analyzed by ANOVA with post-hoc Tukey test (A, D) or unpaired t test (G, H). For non-AUC graphs we used mixed-effects analysis with Tukey’s multiple comparisons. All data are represented as the mean G SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Sections were incubated with polyclonal goat anti-mouse DPP4 (1/40, R&D Systems, AF954), recombinant rabbit monoclonal anti-insulin (1/250; Abcam ab181547, clone number EPR17359), and recombinant anti-glucagon (1/500; Abcam ab92517, clone number EP3070) primary anti- bodies overnight at 4 C. Sections were incubated with Alexa Fluor488 donkey anti-goat (1/500; Thermo Fisher Scientific A32814) and Alexa Fluor Plus 647 donkey anti-rabbit (1/500; Thermo Fisher Scientific A32795) secondary antibodies for 45 minutes and with DAPI for 5 minutes protected from light at room temperature.

Techniques: Derivative Assay, Control, Injection, Isolation

Journal: iScience

Article Title: Pancreas-derived DPP4 is not essential for glucose homeostasis under metabolic stress.

doi: 10.1016/j.isci.2023.106748

Figure Lengend Snippet: Figure 5. Ablation of whole pancreas Dpp4 does not improve glucose tolerance or GSIS (A) Immunofluorescence staining of glucagon (purple) and DPP4 (green) and insulin (purple) and DPP4 (green) in HFD-fed mouse pancreatic sections of PDX- Cre and Dpp4Pan/. DAPI staining (blue) was used to identify nuclei. (B) Absolute mRNA abundance of Dpp4 in islets isolated from one-year-old control (Dpp4(fl/fl)) and Dpp4Pan/ male and female mice. (C and D) Glycemia and AUC glucose during (C) oral and (D) i.p. glucose tolerance tests in HFD-fed control and Dpp4Panl/ male mice. (E) Perifusion GSIS in male mice fed HFD for 25–30 weeks, with and without Exendin 9-39. (F and G) Glycemia and AUC glucose during (F) oral and (G) i.p. glucose tolerance tests in HFD-fed control and Dpp4Pan/ female mice. (H) Perifusion GSIS in female mice fed HFD for 25–30 weeks, with and without Exendin 9-39 (100 nM). Statistical analysis was done with mixed-effects analysis and Tukey’s multiple comparisons (C–H), one-way ANOVA (AUC E and H) or unpaired t test (B, AUC C, D, F and G). All data are represented as the mean G SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Scale bars represent 50 mm.

Article Snippet: Sections were incubated with polyclonal goat anti-mouse DPP4 (1/40, R&D Systems, AF954), recombinant rabbit monoclonal anti-insulin (1/250; Abcam ab181547, clone number EPR17359), and recombinant anti-glucagon (1/500; Abcam ab92517, clone number EP3070) primary anti- bodies overnight at 4 C. Sections were incubated with Alexa Fluor488 donkey anti-goat (1/500; Thermo Fisher Scientific A32814) and Alexa Fluor Plus 647 donkey anti-rabbit (1/500; Thermo Fisher Scientific A32795) secondary antibodies for 45 minutes and with DAPI for 5 minutes protected from light at room temperature.

Techniques: Staining, Isolation, Control

Journal: Medical Sciences

Article Title: Differential Anti-Tumor Effects of IFN-Inducible Chemokines CXCL9, CXCL10, and CXCL11 on a Mouse Squamous Cell Carcinoma Cell Line

doi: 10.3390/medsci11020031

Figure Lengend Snippet: Amino acid sequences of the NH 2 -terminal region of the IFN-inducible chemokine and the cleavage site by DPP4. ( A ). DPP4, also known as CD26, is a serine protease capable of enzymatic removal of the first two amino acids from a protein that possesses proline (P) or alanine (A) in the penultimate NH 2 -terminal position. The first two amino acids of mouse CXCL10 are enzymatically cleaved by DPP4. The truncated form of CXCL10 loses its chemotactic activity and possibly functions as an antagonist that binds to its receptor, CXCR3 [ , ]. ( B ). Amino acid sequence of NH 2 -terminal region of the human IFN-inducible chemokine and the cleavage site by DPP4. These amino acid sequences were obtained from UniProt ( https://www.uniprot.org ; accessed on 25 September 2021).

Article Snippet: The tissue sections were then incubated with goat anti-mouse DPP4 (CD26) polyclonal antibody (AF954; R&D Systems, Minneapolis, MN, USA) at 4 °C in a humidified chamber for 16 h, washed with TBS for 30 min, and blocked with a Histofine Universal Reagent Mouse Stain Kit (Nichirei) at 25 °C for 30 min.

Techniques: Activity Assay, Sequencing

Journal: Medical Sciences

Article Title: Differential Anti-Tumor Effects of IFN-Inducible Chemokines CXCL9, CXCL10, and CXCL11 on a Mouse Squamous Cell Carcinoma Cell Line

doi: 10.3390/medsci11020031

Figure Lengend Snippet: IHC analysis of dipeptidyl peptidase 4 (DPP4) expression in a nude mouse transplanted with chemokine-expressing cells. IHC analysis of DPP4 expression in tumor stroma in nude mice transplanted with chemokine-expressing cells. Original images were captured at 40× ( a , c , e , g , i ): scale bar = 100 μm), and the rectangular regions were captured at 400× in stroma ( b , d , f , h , j ): scale bar = 10 μm).

Article Snippet: The tissue sections were then incubated with goat anti-mouse DPP4 (CD26) polyclonal antibody (AF954; R&D Systems, Minneapolis, MN, USA) at 4 °C in a humidified chamber for 16 h, washed with TBS for 30 min, and blocked with a Histofine Universal Reagent Mouse Stain Kit (Nichirei) at 25 °C for 30 min.

Techniques: Expressing

Journal: Journal of Virology

Article Title: Comparison of Experimental Middle East Respiratory Syndrome Coronavirus Infection Acquired by Three Individual Routes of Infection in the Common Marmoset

doi: 10.1128/jvi.01739-21

Figure Lengend Snippet: Viral replication of MERS-CoV in BHK cells following transfection with either human or marmoset DPP4 receptor RNA. Significant differences were determined by ANOVA, with differences shown between mock-transfected cells (Lipofectamine) and human and marmoset transfected cells (**, P < 0.01; ***, P < 0.001).

Article Snippet: For DPP4 IHC staining, the tissue sections were subjected to heat-induced epitope retrieval using ER1, a citrate-based buffer (catalog number AR9961; Leica Biosystems), for 20 min at 95°C before applying a goat polyclonal anti-human DPP4 antibody (R&D Systems) diluted 1:250 and incubated for 15 min. A rabbit anti-goat secondary IgG antibody (Abcam, UK) was then applied for 8 min before using the Leica Intense R detection kit (Leica Biosystems) for visualization.

Techniques: Transfection

Journal: Journal of Virology

Article Title: Comparison of Experimental Middle East Respiratory Syndrome Coronavirus Infection Acquired by Three Individual Routes of Infection in the Common Marmoset

doi: 10.1128/jvi.01739-21

Figure Lengend Snippet: Location and activity of the DPP4 receptor in marmosets. (A and B) Immunohistochemical staining indicates a very strong presence of the DPP4 receptor within the alveolar spaces of marmosets (A) and no expression in the nasal cavity epithelium and moderate expression in the NALT and other submucosal structures (B). (C) The virus is observed in areas of high DPP4 receptor expression following aerosol challenge with MERS-CoV strain EMC/2012 within the terminal bronchioles and alveolar spaces. (D) An aggregate of MERS-CoV-positive lymphoid cells was observed in the nasal cavity. (E) Locations of MERS-CoV antigen and levels of expression of the DPP4 receptor in the respiratory tract and lungs of marmosets.

Article Snippet: For DPP4 IHC staining, the tissue sections were subjected to heat-induced epitope retrieval using ER1, a citrate-based buffer (catalog number AR9961; Leica Biosystems), for 20 min at 95°C before applying a goat polyclonal anti-human DPP4 antibody (R&D Systems) diluted 1:250 and incubated for 15 min. A rabbit anti-goat secondary IgG antibody (Abcam, UK) was then applied for 8 min before using the Leica Intense R detection kit (Leica Biosystems) for visualization.

Techniques: Activity Assay, Immunohistochemical staining, Staining, Expressing, Virus, Aerosol

Journal: Scientific Reports

Article Title: Insights into replicative senescence of human testicular peritubular cells

doi: 10.1038/s41598-019-51380-w

Figure Lengend Snippet: Proteomic analysis of advanced compared to early passages of HTPCs. Gene set enrichment analysis (GSEA) revealed significantly enriched gene sets (FDR q-value ≤ 0.05) and were summarized using REVIGO by clustering semantically similar GO terms. Each of the 20 characteristic gene sets enriched in early ( a ) and advanced passages ( b ) of HTPCs are shown. Color-coding refers to the corresponding highest GO hierarchy level. The x-axis shows the enrichment significance resulting from the GSEA and is depicted as –log10 (FDR q-value). The number of quantified proteins per gene set is shown in brackets. Volcano plots of intracellular and extracellular proteins, which are more abundant in passaged HTPC cellular proteomes ( c ) and secretomes ( d ) are depicted as red dots and proteins less abundant are shown as blue dots, respectively. Selected proteins with significant difference in abundance are labeled. P-values were calculated by a paired two-sample t -test. DPP4 expression in testicular peritubular cells ( e ). Light micrographs of immunohistochemical staining of human testicular sections. DPP4 is detected in several peritubular cells and cells of the interstitial space. Right micrograph: detail of the DPP4 staining (framed area). The negative control is without staining.

Article Snippet: Primary polyclonal goat anti-human DPP4 antibody (1:40, R&D Systems, Minneapolis, MN, USA) was used.

Techniques: Labeling, Expressing, Immunohistochemical staining, Staining, Negative Control