monoclonal mouse anti mers cov nucleocapsid antibody  (Sino Biological)


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    Name:
    MERS CoV Nucleocapsid Antibody Mouse MAb
    Description:
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant MERS CoV NCoV Novel coronavirus Nucleoprotein NP Catalog 40068 V08B AFS88943 1 Met 1 Asp413 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    Catalog Number:
    40068-MM10
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    MERS CoV
    Applications:
    WB,ELISA
    Immunogen:
    Recombinant MERS-CoV (NCoV / Novel coronavirus) Nucleoprotein / NP protein (Catalog#40068-V08B)
    Product Aliases:
    Anti-coronavirus NP Antibody, Anti-coronavirus Nucleocapsid Antibody, Anti-coronavirus Nucleoprotein Antibody, Anti-cov np Antibody, Anti-ncov NP Antibody, Anti-novel coronavirus Nucleoprotein Antibody, Anti-NP Antibody, Anti-Nucleocapsid Antibody, Anti-Nucleoprotein Antibody
    Antibody Type:
    MAb
    Host:
    Mouse
    Isotype:
    Mouse IgG1
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    Structured Review

    Sino Biological monoclonal mouse anti mers cov nucleocapsid antibody
    Mapping of H2-d restricted T cell epitopes in <t>MERS-CoV</t> N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant MERS CoV NCoV Novel coronavirus Nucleoprotein NP Catalog 40068 V08B AFS88943 1 Met 1 Asp413 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    https://www.bioz.com/result/monoclonal mouse anti mers cov nucleocapsid antibody/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal mouse anti mers cov nucleocapsid antibody - by Bioz Stars, 2021-06
    94/100 stars

    Images

    1) Product Images from "CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice"

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    Journal: Viruses

    doi: 10.3390/v10120718

    Mapping of H2-d restricted T cell epitopes in MERS-CoV N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.
    Figure Legend Snippet: Mapping of H2-d restricted T cell epitopes in MERS-CoV N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.

    Techniques Used: Mouse Assay, Recombinant, Incubation, Enzyme-linked Immunospot

    Identification of an H2-d restricted T cell epitope in MERS-CoV N protein; ( a–d ) Groups of BALB/c mice ( n = 3 to 8) were vaccinated in a prime-boost regime with 10 8 PFU of MVA-MERS-N via i.p. ( a ) or i.m. ( b–d ) application. Mice immunized with non-recombinant MVA (MVA) and PBS served as negative controls. ( a-b ) Splenocytes were stimulated with individual 8-11-mer peptides and IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. ( c–d ) Splenocytes were stimulated with positive MERS-CoV N 10.2 peptide ( c ) or F2L 26-34 peptide ( d ) and IFN-γ producing CD8+ or CD4+ T cells were measured using intracellular cytokine staining assay and FACS analysis. The lines represent means. *
    Figure Legend Snippet: Identification of an H2-d restricted T cell epitope in MERS-CoV N protein; ( a–d ) Groups of BALB/c mice ( n = 3 to 8) were vaccinated in a prime-boost regime with 10 8 PFU of MVA-MERS-N via i.p. ( a ) or i.m. ( b–d ) application. Mice immunized with non-recombinant MVA (MVA) and PBS served as negative controls. ( a-b ) Splenocytes were stimulated with individual 8-11-mer peptides and IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. ( c–d ) Splenocytes were stimulated with positive MERS-CoV N 10.2 peptide ( c ) or F2L 26-34 peptide ( d ) and IFN-γ producing CD8+ or CD4+ T cells were measured using intracellular cytokine staining assay and FACS analysis. The lines represent means. *

    Techniques Used: Mouse Assay, Recombinant, Enzyme-linked Immunospot, Staining, FACS

    Analysis of recombinant MVA-MERS proteins; ( a ) Western Blot analysis of MERS-CoV N protein produced in CEF or HaCat cells. Lysates from cells infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at eight, 12, or 24 hpi. Proteins were analyzed by immunoblotting with a monoclonal anti-MERS-N antibody; ( b – d ) Western Blot analysis of MERS-CoV N and S proteins produced in CEF. Total cell extracts from CEF infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at 24 hpi. Cell lysates and proteins were tested by immunoblotting using monoclonal anti MERS-N and anti MERS-S antibody ( b ) or polyclonal sera from MERS-CoV infected rabbits ( c ) or cynomolgus macaques ( d ). Arrows indicate the N- or S-specific protein bands.
    Figure Legend Snippet: Analysis of recombinant MVA-MERS proteins; ( a ) Western Blot analysis of MERS-CoV N protein produced in CEF or HaCat cells. Lysates from cells infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at eight, 12, or 24 hpi. Proteins were analyzed by immunoblotting with a monoclonal anti-MERS-N antibody; ( b – d ) Western Blot analysis of MERS-CoV N and S proteins produced in CEF. Total cell extracts from CEF infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at 24 hpi. Cell lysates and proteins were tested by immunoblotting using monoclonal anti MERS-N and anti MERS-S antibody ( b ) or polyclonal sera from MERS-CoV infected rabbits ( c ) or cynomolgus macaques ( d ). Arrows indicate the N- or S-specific protein bands.

    Techniques Used: Recombinant, Western Blot, Produced, Infection

    Screening for H2-d restricted T cell epitopes in MERS-CoV N protein using matrix peptide pools; ( a – b ) groups of BALB/c mice ( n = 2 to 6) were vaccinated twice (21-day interval) by i.p. ( a ) or i.m. ( b ) application with 10 8 plaque-forming-units (PFU) of recombinant MVA-MERS-N (MVA-N). Mice inoculated with non-recombinant MVA (MVA) or phosphate-buffered saline (PBS) were used as controls. Splenocytes were restimulated in vitro with pools of overlapping peptides corresponding to MERS-CoV N protein. IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. The lines represent means.
    Figure Legend Snippet: Screening for H2-d restricted T cell epitopes in MERS-CoV N protein using matrix peptide pools; ( a – b ) groups of BALB/c mice ( n = 2 to 6) were vaccinated twice (21-day interval) by i.p. ( a ) or i.m. ( b ) application with 10 8 plaque-forming-units (PFU) of recombinant MVA-MERS-N (MVA-N). Mice inoculated with non-recombinant MVA (MVA) or phosphate-buffered saline (PBS) were used as controls. Splenocytes were restimulated in vitro with pools of overlapping peptides corresponding to MERS-CoV N protein. IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. The lines represent means.

    Techniques Used: Mouse Assay, Recombinant, In Vitro, Enzyme-linked Immunospot

    Related Articles

    Incubation:

    Article Title: Therapeutic effect of CT-P59 against SARS-CoV-2 South African variant
    Article Snippet: .. The cells were formalin-fixed and ethanol permeabilized followed by incubation with a murine monoclonal antibody which targets the viral nucleocapsid protein (Sino Biological), followed by a secondary anti-mouse IgG peroxidase conjugate (Thermo Scientific) and TrueBlue (KPL) substrate. ..

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice
    Article Snippet: Total cell proteins were resolved by electrophoresis in a sodium dodecyl sulfate (SDS)-10% polyacrylamide gel (SDS-PAGE) and subsequently transferred onto a nitrocellulose membrane via electroblotting. .. After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies. .. After washing with 0.1% NP-40 in PBS, the blots were incubated with anti-mouse IgG (1:5000), or anti-rabbit IgG antibody (1:5000), or protein A (1:1000) conjugated to horseradish peroxidase (Cell Signaling Technology, Frankfurt am Main, Germany).

    Blocking Assay:

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice
    Article Snippet: Total cell proteins were resolved by electrophoresis in a sodium dodecyl sulfate (SDS)-10% polyacrylamide gel (SDS-PAGE) and subsequently transferred onto a nitrocellulose membrane via electroblotting. .. After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies. .. After washing with 0.1% NP-40 in PBS, the blots were incubated with anti-mouse IgG (1:5000), or anti-rabbit IgG antibody (1:5000), or protein A (1:1000) conjugated to horseradish peroxidase (Cell Signaling Technology, Frankfurt am Main, Germany).

    Infection:

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice
    Article Snippet: Total cell proteins were resolved by electrophoresis in a sodium dodecyl sulfate (SDS)-10% polyacrylamide gel (SDS-PAGE) and subsequently transferred onto a nitrocellulose membrane via electroblotting. .. After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies. .. After washing with 0.1% NP-40 in PBS, the blots were incubated with anti-mouse IgG (1:5000), or anti-rabbit IgG antibody (1:5000), or protein A (1:1000) conjugated to horseradish peroxidase (Cell Signaling Technology, Frankfurt am Main, Germany).

    Article Title: Blocking transmission of Middle East respiratory syndrome coronavirus (MERS-CoV) in llamas by vaccination with a recombinant spike protein
    Article Snippet: .. After 8 h of infection, the cells were fixed and stained using mouse anti-MERS-CoV nucleocapsid protein (SinoBiological) and HRP-conjugated goat anti-mouse IgG1 (SouthernBiotech). .. The number of infected cells were detected using a precipitate-forming TMB substrate (True Blue, KPL) and counted using an ImmunoSpot® Image analyser (CTL Europe GmbH).

    Staining:

    Article Title: Blocking transmission of Middle East respiratory syndrome coronavirus (MERS-CoV) in llamas by vaccination with a recombinant spike protein
    Article Snippet: .. After 8 h of infection, the cells were fixed and stained using mouse anti-MERS-CoV nucleocapsid protein (SinoBiological) and HRP-conjugated goat anti-mouse IgG1 (SouthernBiotech). .. The number of infected cells were detected using a precipitate-forming TMB substrate (True Blue, KPL) and counted using an ImmunoSpot® Image analyser (CTL Europe GmbH).

    Expressing:

    Article Title: Species-Specific Colocalization of Middle East Respiratory Syndrome Coronavirus Attachment and Entry Receptors
    Article Snippet: The bat tissues included in this study were histologically normal as determined by using hematoxylin-eosin staining prior to our experiment. .. MERS-CoV nucleoprotein was detected with 5 μg/ml mouse anti-MERS nucleoprotein (Sino-Biological, Beijing, China), while DPP4 expression was detected with either 5 μg/ml goat anti-human DPP4 (R & D, Minneapolis, MN, USA) or 10 μg/ml mouse anti-human DPP4 (clone 11D7; Origene, Rockville, MD, USA). .. Briefly, the paraffin-embedded tissues were deparaffinized using xylene, hydrated using graded concentrations of alcohol, boiled in 10 mM citric acid buffer (pH 6) for 15 min, and subsequently incubated in 3% H2 O2 for 10 min and in 5% normal goat serum for 30 min before staining with antibodies.

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    Sino Biological monoclonal mouse anti mers cov nucleocapsid antibody
    Mapping of H2-d restricted T cell epitopes in <t>MERS-CoV</t> N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.
    Monoclonal Mouse Anti Mers Cov Nucleocapsid Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal mouse anti mers cov nucleocapsid antibody/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal mouse anti mers cov nucleocapsid antibody - by Bioz Stars, 2021-06
    94/100 stars
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    Mapping of H2-d restricted T cell epitopes in MERS-CoV N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Mapping of H2-d restricted T cell epitopes in MERS-CoV N protein; ( a–b ) BALB/c mice (n = 2 to 4) were immunized twice (21-day interval) i.p. or i.m. with 10 8 PFU of recombinant MVA-MERS-N (MVA-N), non-recombinant MVA (MVA) or PBS. Splenocytes from vaccinated mice were incubated in the presence of subpools (V8.1, V8.2, H8.1, H8.2) from positive matrix pools ( a ) or individual 15-mers peptides #89 or #90 ( b ). IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were quantified by ELISPOT. The lines represent means.

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Mouse Assay, Recombinant, Incubation, Enzyme-linked Immunospot

    Identification of an H2-d restricted T cell epitope in MERS-CoV N protein; ( a–d ) Groups of BALB/c mice ( n = 3 to 8) were vaccinated in a prime-boost regime with 10 8 PFU of MVA-MERS-N via i.p. ( a ) or i.m. ( b–d ) application. Mice immunized with non-recombinant MVA (MVA) and PBS served as negative controls. ( a-b ) Splenocytes were stimulated with individual 8-11-mer peptides and IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. ( c–d ) Splenocytes were stimulated with positive MERS-CoV N 10.2 peptide ( c ) or F2L 26-34 peptide ( d ) and IFN-γ producing CD8+ or CD4+ T cells were measured using intracellular cytokine staining assay and FACS analysis. The lines represent means. *

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Identification of an H2-d restricted T cell epitope in MERS-CoV N protein; ( a–d ) Groups of BALB/c mice ( n = 3 to 8) were vaccinated in a prime-boost regime with 10 8 PFU of MVA-MERS-N via i.p. ( a ) or i.m. ( b–d ) application. Mice immunized with non-recombinant MVA (MVA) and PBS served as negative controls. ( a-b ) Splenocytes were stimulated with individual 8-11-mer peptides and IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. ( c–d ) Splenocytes were stimulated with positive MERS-CoV N 10.2 peptide ( c ) or F2L 26-34 peptide ( d ) and IFN-γ producing CD8+ or CD4+ T cells were measured using intracellular cytokine staining assay and FACS analysis. The lines represent means. *

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Mouse Assay, Recombinant, Enzyme-linked Immunospot, Staining, FACS

    Analysis of recombinant MVA-MERS proteins; ( a ) Western Blot analysis of MERS-CoV N protein produced in CEF or HaCat cells. Lysates from cells infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at eight, 12, or 24 hpi. Proteins were analyzed by immunoblotting with a monoclonal anti-MERS-N antibody; ( b – d ) Western Blot analysis of MERS-CoV N and S proteins produced in CEF. Total cell extracts from CEF infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at 24 hpi. Cell lysates and proteins were tested by immunoblotting using monoclonal anti MERS-N and anti MERS-S antibody ( b ) or polyclonal sera from MERS-CoV infected rabbits ( c ) or cynomolgus macaques ( d ). Arrows indicate the N- or S-specific protein bands.

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Analysis of recombinant MVA-MERS proteins; ( a ) Western Blot analysis of MERS-CoV N protein produced in CEF or HaCat cells. Lysates from cells infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at eight, 12, or 24 hpi. Proteins were analyzed by immunoblotting with a monoclonal anti-MERS-N antibody; ( b – d ) Western Blot analysis of MERS-CoV N and S proteins produced in CEF. Total cell extracts from CEF infected with recombinant MVA (MVA-MERS-N, MVA-MERS-S) or non-recombinant MVA (MVA) at a MOI of five, or from non-infected cells (mock) were prepared at 24 hpi. Cell lysates and proteins were tested by immunoblotting using monoclonal anti MERS-N and anti MERS-S antibody ( b ) or polyclonal sera from MERS-CoV infected rabbits ( c ) or cynomolgus macaques ( d ). Arrows indicate the N- or S-specific protein bands.

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Recombinant, Western Blot, Produced, Infection

    Screening for H2-d restricted T cell epitopes in MERS-CoV N protein using matrix peptide pools; ( a – b ) groups of BALB/c mice ( n = 2 to 6) were vaccinated twice (21-day interval) by i.p. ( a ) or i.m. ( b ) application with 10 8 plaque-forming-units (PFU) of recombinant MVA-MERS-N (MVA-N). Mice inoculated with non-recombinant MVA (MVA) or phosphate-buffered saline (PBS) were used as controls. Splenocytes were restimulated in vitro with pools of overlapping peptides corresponding to MERS-CoV N protein. IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. The lines represent means.

    Journal: Viruses

    Article Title: CD8+ T Cells Responding to the Middle East Respiratory Syndrome Coronavirus Nucleocapsid Protein Delivered by Vaccinia Virus MVA in Mice

    doi: 10.3390/v10120718

    Figure Lengend Snippet: Screening for H2-d restricted T cell epitopes in MERS-CoV N protein using matrix peptide pools; ( a – b ) groups of BALB/c mice ( n = 2 to 6) were vaccinated twice (21-day interval) by i.p. ( a ) or i.m. ( b ) application with 10 8 plaque-forming-units (PFU) of recombinant MVA-MERS-N (MVA-N). Mice inoculated with non-recombinant MVA (MVA) or phosphate-buffered saline (PBS) were used as controls. Splenocytes were restimulated in vitro with pools of overlapping peptides corresponding to MERS-CoV N protein. IFN-γ spot-forming CD8+ T cells (IFN-γ SFC) were measured by ELISPOT. The lines represent means.

    Article Snippet: After 1 h blocking in a phosphate buffered saline (PBS) buffer containing 1% (w/v) non-fat dried milk and 0.1% (v/v) NP-40 detergent, the blots were incubated with monoclonal mouse anti-MERS-CoV Nucleocapsid antibody (Sino Biological, Beijing, China, 1:1000), monoclonal rabbit anti-MERS-CoV Spike Protein S1 Antibody (Sino Biological, 1:500), or polyclonal sera from MERS-CoV infected rabbits or cynomolgus macaques (kindly provided by Dr. Bart Haagmans, Erasmus Medical Center, Rotterdam, 1:1000) [ ] as primary antibodies.

    Techniques: Mouse Assay, Recombinant, In Vitro, Enzyme-linked Immunospot

    DPP4 expression and MERS-CoV S1 A binding in intestinal tissues of common pipistrelle bat, serotine bat, Gambian epauletted fruit bat, and Egyptian fruit bat. DPP4 expression and MERS-CoV S1 A binding are indicated in red. DPP4 is expressed at the apical surface of the intestinal epithelial cells of these four bat species. MERS-CoV S1 A binds to the apical surface of the intestinal epithelial cells of common pipistrelle bats in both villi and crypts, while in other bat species, it mostly binds to intestinal epithelial cells within the crypts. Magnification, ×400.

    Journal: Journal of Virology

    Article Title: Species-Specific Colocalization of Middle East Respiratory Syndrome Coronavirus Attachment and Entry Receptors

    doi: 10.1128/JVI.00107-19

    Figure Lengend Snippet: DPP4 expression and MERS-CoV S1 A binding in intestinal tissues of common pipistrelle bat, serotine bat, Gambian epauletted fruit bat, and Egyptian fruit bat. DPP4 expression and MERS-CoV S1 A binding are indicated in red. DPP4 is expressed at the apical surface of the intestinal epithelial cells of these four bat species. MERS-CoV S1 A binds to the apical surface of the intestinal epithelial cells of common pipistrelle bats in both villi and crypts, while in other bat species, it mostly binds to intestinal epithelial cells within the crypts. Magnification, ×400.

    Article Snippet: MERS-CoV nucleoprotein was detected with 5 μg/ml mouse anti-MERS nucleoprotein (Sino-Biological, Beijing, China), while DPP4 expression was detected with either 5 μg/ml goat anti-human DPP4 (R & D, Minneapolis, MN, USA) or 10 μg/ml mouse anti-human DPP4 (clone 11D7; Origene, Rockville, MD, USA).

    Techniques: Expressing, Binding Assay

    MERS-CoV S1 A binds specifically to the nasal epithelium of dromedary camels. (A) Nanoparticles displaying a multivalent MERS-CoV S1 A domain (np-S1 A ) bind to the apical surface of camel nasal ciliated epithelial cells, as revealed by red staining. np-S1 A binding is inhibited by prior neuraminidase treatment of these nasal tissues. Blank nanoparticles also do not bind to these tissues. (B) Goblet cells (arrows), visualized in purple by periodic acid-Schiff stain, are DPP4 negative, unlike nasal ciliated columnar epithelial cells (arrowheads). DPP4 expression is indicated in red. MERS-CoV S1 A binding to these goblet cells (red) can be abrogated by neuraminidase treatment. In MERS-CoV-infected camels, MERS-CoV S1 A (green) binds to both nasal ciliated columnar epithelial cells and goblet cells, while MERS-CoV N protein (red) is detected only in nasal ciliated columnar epithelial cells. np-S1 A binding is abrogated by a nanobody against the S1 A domain (Nb anti-S1 A ) but not by one against the S1 B domain (Nb anti-S1 B ). The tissues used in these experiments were sequentially cut. All pictures were taken at a ×400 magnification. (C) Nb anti-S1 A and Nb anti-S1 B bind specifically to S1 A and S1 B domains, respectively, and both bind to S1 protein, as revealed by an ELISA. The control nanobody does not bind to S1, S1 A , and S1 B . Nanobody binding is expressed as optical density at 450 nm (OD 450 ) values determined by an ELISA.

    Journal: Journal of Virology

    Article Title: Species-Specific Colocalization of Middle East Respiratory Syndrome Coronavirus Attachment and Entry Receptors

    doi: 10.1128/JVI.00107-19

    Figure Lengend Snippet: MERS-CoV S1 A binds specifically to the nasal epithelium of dromedary camels. (A) Nanoparticles displaying a multivalent MERS-CoV S1 A domain (np-S1 A ) bind to the apical surface of camel nasal ciliated epithelial cells, as revealed by red staining. np-S1 A binding is inhibited by prior neuraminidase treatment of these nasal tissues. Blank nanoparticles also do not bind to these tissues. (B) Goblet cells (arrows), visualized in purple by periodic acid-Schiff stain, are DPP4 negative, unlike nasal ciliated columnar epithelial cells (arrowheads). DPP4 expression is indicated in red. MERS-CoV S1 A binding to these goblet cells (red) can be abrogated by neuraminidase treatment. In MERS-CoV-infected camels, MERS-CoV S1 A (green) binds to both nasal ciliated columnar epithelial cells and goblet cells, while MERS-CoV N protein (red) is detected only in nasal ciliated columnar epithelial cells. np-S1 A binding is abrogated by a nanobody against the S1 A domain (Nb anti-S1 A ) but not by one against the S1 B domain (Nb anti-S1 B ). The tissues used in these experiments were sequentially cut. All pictures were taken at a ×400 magnification. (C) Nb anti-S1 A and Nb anti-S1 B bind specifically to S1 A and S1 B domains, respectively, and both bind to S1 protein, as revealed by an ELISA. The control nanobody does not bind to S1, S1 A , and S1 B . Nanobody binding is expressed as optical density at 450 nm (OD 450 ) values determined by an ELISA.

    Article Snippet: MERS-CoV nucleoprotein was detected with 5 μg/ml mouse anti-MERS nucleoprotein (Sino-Biological, Beijing, China), while DPP4 expression was detected with either 5 μg/ml goat anti-human DPP4 (R & D, Minneapolis, MN, USA) or 10 μg/ml mouse anti-human DPP4 (clone 11D7; Origene, Rockville, MD, USA).

    Techniques: Staining, Binding Assay, Expressing, Infection, Enzyme-linked Immunosorbent Assay

    MERS-CoV receptor and attachment factor in the human lower respiratory tract epithelium. (A) The MERS-CoV receptor, DPP4, is expressed in the nasal epithelium of dromedary camels, while in the lungs, it is mainly expressed in endothelial cells. In the human respiratory tract, DPP4 is expressed in bronchiolar epithelial cells (arrowhead) and type II pneumocytes (arrow) in the lungs but not in the nasal epithelium. (B) α2,3-Sialic acid expression and MERS-CoV S1 A binding are also detected in human bronchiolar epithelial cells (arrowheads) and type II pneumocytes (arrows). DPP4 expression, α2,3-sialic acids, and MERS-CoV S1 A binding are indicated in red. (C) In human alveoli, DPP4 expression (red) colocalizes in the same cells where MERS-CoV S1 A binds (green). Pictures of the nasal epithelium were taken at a ×400 magnification, and those of the alveoli were taken at a ×1,000 magnification.

    Journal: Journal of Virology

    Article Title: Species-Specific Colocalization of Middle East Respiratory Syndrome Coronavirus Attachment and Entry Receptors

    doi: 10.1128/JVI.00107-19

    Figure Lengend Snippet: MERS-CoV receptor and attachment factor in the human lower respiratory tract epithelium. (A) The MERS-CoV receptor, DPP4, is expressed in the nasal epithelium of dromedary camels, while in the lungs, it is mainly expressed in endothelial cells. In the human respiratory tract, DPP4 is expressed in bronchiolar epithelial cells (arrowhead) and type II pneumocytes (arrow) in the lungs but not in the nasal epithelium. (B) α2,3-Sialic acid expression and MERS-CoV S1 A binding are also detected in human bronchiolar epithelial cells (arrowheads) and type II pneumocytes (arrows). DPP4 expression, α2,3-sialic acids, and MERS-CoV S1 A binding are indicated in red. (C) In human alveoli, DPP4 expression (red) colocalizes in the same cells where MERS-CoV S1 A binds (green). Pictures of the nasal epithelium were taken at a ×400 magnification, and those of the alveoli were taken at a ×1,000 magnification.

    Article Snippet: MERS-CoV nucleoprotein was detected with 5 μg/ml mouse anti-MERS nucleoprotein (Sino-Biological, Beijing, China), while DPP4 expression was detected with either 5 μg/ml goat anti-human DPP4 (R & D, Minneapolis, MN, USA) or 10 μg/ml mouse anti-human DPP4 (clone 11D7; Origene, Rockville, MD, USA).

    Techniques: Expressing, Binding Assay

    Detection of MERS-CoV N protein, DPP4, α2,3-sialic acids, and MERS-CoV S1 A binding in the nasal epithelium of dromedary camels, pigs, and rabbits. MERS-CoV N protein, DPP4, α2,3-sialic acid, and MERS-CoV S1 A binding are all indicated in red. MERS-CoV N protein is detected in the nasal epithelium tissues of MERS-CoV-infected dromedary camels, pigs, and rabbits. DPP4, α2,3-sialic acid, and MERS-CoV S1 A binding were evaluated on the tissues of noninfected animals. MERS-CoV N protein and DPP4 are detected in the nasal epithelium of dromedary camel, pig, and rabbit. α2,3-Sialic acids are detected in the nasal epithelium of dromedary camel and rabbit but not in that of pig. Meanwhile, MERS-CoV S1 A binds merely to the nasal epithelium of dromedary camel. Magnification, ×400.

    Journal: Journal of Virology

    Article Title: Species-Specific Colocalization of Middle East Respiratory Syndrome Coronavirus Attachment and Entry Receptors

    doi: 10.1128/JVI.00107-19

    Figure Lengend Snippet: Detection of MERS-CoV N protein, DPP4, α2,3-sialic acids, and MERS-CoV S1 A binding in the nasal epithelium of dromedary camels, pigs, and rabbits. MERS-CoV N protein, DPP4, α2,3-sialic acid, and MERS-CoV S1 A binding are all indicated in red. MERS-CoV N protein is detected in the nasal epithelium tissues of MERS-CoV-infected dromedary camels, pigs, and rabbits. DPP4, α2,3-sialic acid, and MERS-CoV S1 A binding were evaluated on the tissues of noninfected animals. MERS-CoV N protein and DPP4 are detected in the nasal epithelium of dromedary camel, pig, and rabbit. α2,3-Sialic acids are detected in the nasal epithelium of dromedary camel and rabbit but not in that of pig. Meanwhile, MERS-CoV S1 A binds merely to the nasal epithelium of dromedary camel. Magnification, ×400.

    Article Snippet: MERS-CoV nucleoprotein was detected with 5 μg/ml mouse anti-MERS nucleoprotein (Sino-Biological, Beijing, China), while DPP4 expression was detected with either 5 μg/ml goat anti-human DPP4 (R & D, Minneapolis, MN, USA) or 10 μg/ml mouse anti-human DPP4 (clone 11D7; Origene, Rockville, MD, USA).

    Techniques: Binding Assay, Infection

    Binding of the MERS-CoV S1 A domain and MERS-CoV infection in primary normal human bronchial epithelial cells are inhibited upon prior neuraminidase treatment. (A) Removal of sialic acids using neuraminidase (NA) treatment diminishes MERS-CoV S1 A binding to primary normal human bronchial epithelial cells but not DPP4 expression in these cells. (B) The same treatment also significantly inhibits MERS-CoV infection in these cells up to 50%. Immunofluorescence images in panel A were taken at a ×400 magnification, and those in panel B were taken at a ×100 magnification. ***, P value of

    Journal: Journal of Virology

    Article Title: Species-Specific Colocalization of Middle East Respiratory Syndrome Coronavirus Attachment and Entry Receptors

    doi: 10.1128/JVI.00107-19

    Figure Lengend Snippet: Binding of the MERS-CoV S1 A domain and MERS-CoV infection in primary normal human bronchial epithelial cells are inhibited upon prior neuraminidase treatment. (A) Removal of sialic acids using neuraminidase (NA) treatment diminishes MERS-CoV S1 A binding to primary normal human bronchial epithelial cells but not DPP4 expression in these cells. (B) The same treatment also significantly inhibits MERS-CoV infection in these cells up to 50%. Immunofluorescence images in panel A were taken at a ×400 magnification, and those in panel B were taken at a ×100 magnification. ***, P value of

    Article Snippet: MERS-CoV nucleoprotein was detected with 5 μg/ml mouse anti-MERS nucleoprotein (Sino-Biological, Beijing, China), while DPP4 expression was detected with either 5 μg/ml goat anti-human DPP4 (R & D, Minneapolis, MN, USA) or 10 μg/ml mouse anti-human DPP4 (clone 11D7; Origene, Rockville, MD, USA).

    Techniques: Binding Assay, Infection, Expressing, Immunofluorescence

    Experimental infection of TMPRSS2-knockout hDPP4-transgenic (TMPRSS2-KO Tg) mice with MERS-CoV. hDPP4-Tg (Tg) and TMPRSS2-KO Tg (KO-Tg) mice were inoculated with EMC-HCoV (MERS-CoV). (a) Body weight curve during the 14 days postinfection (p.i.). Numbers of animals per group were as follows: TMPRSS2-KO Tg, n = 8 (male, 2; female, 6); hDPP4-Tg, n = 6 (male, 3; female, 3). Mice 12 to 14 weeks old were used. Error bars represent standard errors (***, P

    Journal: Journal of Virology

    Article Title: TMPRSS2 Contributes to Virus Spread and Immunopathology in the Airways of Murine Models after Coronavirus Infection

    doi: 10.1128/JVI.01815-18

    Figure Lengend Snippet: Experimental infection of TMPRSS2-knockout hDPP4-transgenic (TMPRSS2-KO Tg) mice with MERS-CoV. hDPP4-Tg (Tg) and TMPRSS2-KO Tg (KO-Tg) mice were inoculated with EMC-HCoV (MERS-CoV). (a) Body weight curve during the 14 days postinfection (p.i.). Numbers of animals per group were as follows: TMPRSS2-KO Tg, n = 8 (male, 2; female, 6); hDPP4-Tg, n = 6 (male, 3; female, 3). Mice 12 to 14 weeks old were used. Error bars represent standard errors (***, P

    Article Snippet: Antigen retrieval from formalin-fixed mouse tissue sections was performed by autoclaving in retrieval solution (pH 6.0) (Nichirei Biosciences) at 121°C for 10 min. Hyperimmune rabbit serum raised against SARS-CoV ( ) or an anti-MERS-CoV nucleocapsid antibody (Sino Biological Inc., Beijing, China) was used as the primary antibody to detect viral antigens.

    Techniques: Infection, Knock-Out, Transgenic Assay, Mouse Assay