goat polyclonal anti human dpp4 antibody  (R&D Systems)

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

Journal: Emerging Infectious Diseases

Article Title: Livestock Susceptibility to Infection with Middle East Respiratory Syndrome Coronavirus

doi: 10.3201/eid2302.161239

Figure Lengend Snippet: Presence of MERS-CoV receptor DPP4 (IHC) and of mucosubstances (PAS) in upper and lower respiratory tract tissues from sheep, pigs, llamas, and horses. A) In the nose, DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of pigs, llamas, and horses but not sheep. PAS staining (magenta) demonstrated more mucous cells in the lining epithelium of sheep and horses and a layer of mucus on the lining epithelium of the horses. B) DPP4 (red cytoplasmic or membrane staining) was present on the lining epithelium of the trachea, bronchus/bronchioles, and alveoli in the pigs, llamas and horses but not in the sheep. Original magnification ×400 for all images. DPP4, dipeptidyl peptidase-4; IHC, immunohistochemistry; MERS-CoV, Middle East respiratory syndrome coronavirus; PAS, periodic acid–Schiff; term., terminal.

Article Snippet: In brief, we performed DPP4 IHC staining by using 5 μg/mL of polyclonal goat IgG anti-human DPP4 antibody (R&D Systems, Abingdon, UK) and peroxidase-labeled rabbit anti-goat IgG (1:200; DAKO; Agilent Technologies Company, Santa Clara, CA, USA) as a secondary antibody.

Techniques: Membrane, Staining, Immunohistochemistry

Journal: Journal of Virology

Article Title: Differential Expression of the Middle East Respiratory Syndrome Coronavirus Receptor in the Upper Respiratory Tracts of Humans and Dromedary Camels

doi: 10.1128/jvi.02994-15

Figure Lengend Snippet: Figure 1. DPP4 expression in the upper respiratory tract of camels and humans. DPP4 293

Article Snippet: DPP4 was detected using 5 μg/ml polyclonal goat IgG anti-human DPP4 antibody 82 (R&D systems, Abingdon, UK), while negative controls were stained using normal goat serum 83 (MP Biomedicals, Santa Ana, CA, USA) in equal concentration.

Techniques: Expressing

Journal: Archives of Virology

Article Title: Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein

doi: 10.1007/s00705-015-2506-z

Figure Lengend Snippet: Rat DPP4 does not act as a MERS-CoV receptor. (A) The expression of DPP4 on CHO cells. CHO cells were transfected with expression plasmids encoding human and rat DPP4. Empty pcDNA was used as a negative control. The expression of DPP4 was detected using an anti-human DPP4 polyclonal antibody, followed by a FITC-labeled anti-goat antibody. (B) Cells transfected with expression plasmids encoding human or rat DPP4 or the pcDNA vector were inoculated with MERSpv-St16 (MOI = 0.2). GFP expression was examined by fluorescence microscopy. (C) Cells transfected with expression plasmids encoding human or rat DPP4 or the pcDNA vector were inoculated with MERS-CoV (MOI = 0.1). At 48 hours postinfection, the cells were stained with anti-MERS N rabbit serum

Article Snippet: Cells transfected with pcDNA bearing human or rat DPP4 were fixed with 10 % formalin under non-permeabilizing conditions and stained with polyclonal goat anti-human DPP4 (R&D Systems, Minneapolis, MN).

Techniques: Expressing, Transfection, Negative Control, Labeling, Plasmid Preparation, Fluorescence, Microscopy, Staining

Journal: Archives of Virology

Article Title: Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein

doi: 10.1007/s00705-015-2506-z

Figure Lengend Snippet: DPP4 residues in contact with RBD of MERS-CoV

Article Snippet: Cells transfected with pcDNA bearing human or rat DPP4 were fixed with 10 % formalin under non-permeabilizing conditions and stained with polyclonal goat anti-human DPP4 (R&D Systems, Minneapolis, MN).

Techniques:

Journal: Archives of Virology

Article Title: Inability of rat DPP4 to allow MERS-CoV infection revealed by using a VSV pseudotype bearing truncated MERS-CoV spike protein

doi: 10.1007/s00705-015-2506-z

Figure Lengend Snippet: Mutational analysis of rat DPP4. CHO cells transfected with expression plasmids encoding human DPP4, rat DPP4, or rat DPP4 mutants were inoculated with MERSpv-St16. GFP expression was examined by fluorescence microscopy, and the number of the GFP-positive cells was determined

Article Snippet: Cells transfected with pcDNA bearing human or rat DPP4 were fixed with 10 % formalin under non-permeabilizing conditions and stained with polyclonal goat anti-human DPP4 (R&D Systems, Minneapolis, MN).

Techniques: Transfection, Expressing, Fluorescence, Microscopy

Journal: Nature

Article Title: Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC

doi: 10.1038/nature12005

Figure Lengend Snippet: a , Huh-7 cell lysates were incubated with hCoV-EMC and SARS-CoV S1–Fc proteins and affinity-isolated proteins were subjected to protein electrophoresis under non-reducing conditions. The arrowhead indicates the position of the ∼110-kDa DPP4 protein specifically isolated using the hCoV-EMC S1–Fc protein. b , hCoV-EMC and SARS-CoV S1–Fc proteins were mock-incubated or incubated with soluble DPP4 (sDPP4) or soluble ACE2 (sACE2) followed by protein A sepharose affinity isolation and subjected to protein electrophoresis under non-reducing conditions.

Article Snippet: After 24 h incubation, cells were stained with both goat-anti-human DPP4 polyclonal antibody (R&D system) and a rabbit anti-goat IgG–FITC antibody (Sigma).

Techniques: Incubation, Isolation, Protein Electrophoresis

Journal: Nature

Article Title: Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC

doi: 10.1038/nature12005

Figure Lengend Snippet: a , COS-7 cells transfected with plasmids encoding human DPP4 (hDPP4), bat DPP4 (bDPP4) or a control plasmid (pcDNA) were tested for S1 binding and staining with a polyclonal antiserum against DPP4. b , Similarly, COS-7, Huh-7, Vero and bat cells were tested for reactivity with the same antiserum against DPP4 (blue lines) or with a control normal goat serum (grey peak). c , d , DPP4 expression (red) was also found in primary human bronchiolar epithelial cell cultures ( c ) and human bronchiolar tissue ( d ) and appeared to be localized to the apical surfaces of non-ciliated cells that do not express β-tubulin IV (green). e , Double-stranded viral RNA (cyan) was detected in hCoV-EMC-infected primary human bronchiolar epithelial cell cultures and appeared to be localized to non-ciliated cells that express DPP4 (red). Stainings were performed using antibodies directed against β-tubulin IV (ciliated cells; green), DPP4 (red), dsRNA (hCoV-EMC; cyan), and DAPI (cell nucleus; blue). All scale bars are 10 μm.

Article Snippet: After 24 h incubation, cells were stained with both goat-anti-human DPP4 polyclonal antibody (R&D system) and a rabbit anti-goat IgG–FITC antibody (Sigma).

Techniques: Transfection, Plasmid Preparation, Binding Assay, Staining, Expressing, Infection

Journal: Nature

Article Title: Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC

doi: 10.1038/nature12005

Figure Lengend Snippet: a , Inhibition of hCoV-EMC infection of Huh-7 cells by antibodies to DPP4. Supernatants collected at 2 h (open bars) and 20 h (filled bars) were tested for the presence of hCoV-EMC RNA using a TaqMan assay. Results representative of three different experiments are shown as ΔCt values (one-way ANOVA test, * P < 0.05; n = 3 per group), normal goat, normal goat serum. b , Infection of human primary bronchiolar epithelial cells is blocked by the DPP4 antibodies in a dose-dependent manner and samples were analysed at 2 h (open bars) and 20 h (filled bars) after infection (one-way ANOVA test, * P < 0.05; n = 3 per group). c , COS-7 cells transfected with plasmids encoding human DPP4 (hDPP4), bat DPP4 (bDPP4) or a control plasmid (pcDNA) were inoculated with hCoV-EMC at a multiplicity of infection of 1 and left for 1 h. Cells were washed twice and stained at 8 h after infection (original magnification, ×200) or supernatant collected at 2 h (open bars), 20 h (black bars) and 40 h (blue bars) was tested for the presence of hCoV-EMC RNA using a TaqMan assay ( d ). Results representative of three different experiments are expressed as GE (TCID 50 ml −1 ) values (one-way ANOVA test, * P < 0.05; n = 4 per group). All error bars represent s.e.m.

Article Snippet: After 24 h incubation, cells were stained with both goat-anti-human DPP4 polyclonal antibody (R&D system) and a rabbit anti-goat IgG–FITC antibody (Sigma).

Techniques: Inhibition, Infection, TaqMan Assay, Transfection, Plasmid Preparation, Staining