mers cov nucleoprotein np antibody rabbit pab  (Sino Biological)


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    Name:
    MERS CoV Nucleoprotein NP Antibody Rabbit PAb
    Description:
    Produced in rabbits immunized with purified recombinant MERS CoV NCoV Novel coronavirus Nucleoprotein NP Catalog 40068 V08B AFS88943 1 Met1 Asp413 Total IgG was purified by Protein A affinity chromatography
    Catalog Number:
    40068-RP01
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    MERS CoV
    Applications:
    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:
    PAb
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Sino Biological mers cov nucleoprotein np antibody rabbit pab
    ΔORF5 <t>MERS-CoV</t> infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P
    Produced in rabbits immunized with purified recombinant MERS CoV NCoV Novel coronavirus Nucleoprotein NP Catalog 40068 V08B AFS88943 1 Met1 Asp413 Total IgG was purified by Protein A affinity chromatography
    https://www.bioz.com/result/mers cov nucleoprotein np antibody rabbit pab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers cov nucleoprotein np antibody rabbit pab - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus"

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-64264-1

    ΔORF5 MERS-CoV infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P
    Figure Legend Snippet: ΔORF5 MERS-CoV infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P

    Techniques Used: Infection

    MERS-CoV gene expression varies between acute and persistently infected bat cells. RNA from persistently infected Efk cells and acutely infected Efk cells were harvested at several time points and MERS-CoV genome quantities (upE levels) and gene expression levels were analyzed by real time quantitative PCR. ( a–i) Genome and gene expression (−(ΔCT gene − ΔCT GAPDH )) levels for MERS-CoV upE, S, ORF3, ORF4a, ORF4b, ORF5, E, M and N genes in acute (green) vs. persistent (purple) infections at 0, 12, 24 and 48 hours post-infection or seeding, respectively (n = 4; Mean ± SD). ***P
    Figure Legend Snippet: MERS-CoV gene expression varies between acute and persistently infected bat cells. RNA from persistently infected Efk cells and acutely infected Efk cells were harvested at several time points and MERS-CoV genome quantities (upE levels) and gene expression levels were analyzed by real time quantitative PCR. ( a–i) Genome and gene expression (−(ΔCT gene − ΔCT GAPDH )) levels for MERS-CoV upE, S, ORF3, ORF4a, ORF4b, ORF5, E, M and N genes in acute (green) vs. persistent (purple) infections at 0, 12, 24 and 48 hours post-infection or seeding, respectively (n = 4; Mean ± SD). ***P

    Techniques Used: Expressing, Infection, Real-time Polymerase Chain Reaction

    Proposed model for establishment of persistent MERS-CoV infection in bat cells. As with the stocks of most RNA viruses, the MERS-CoV inoculum is made up of the dominant W + virus as well as smaller numbers of variants, including variants with inactivating mutations in ORF5 (ΔORF5). The cells infected with the more cytolytic W + virus die, while the small number of cells infected with ΔORF5 MERS-CoV survive because of an ensuing antiviral response and the induction of anti-apoptotic processes. ΔORF5 MERS-CoV infected cells are resistant to infection with the W + virus and the W + virus is soon diluted out. After a process of cell death and recovery, ΔORF5 MERS-CoV infected cells survive and take over, leading to a culture of persistently infected cells that produce small, but consistent amounts of virus over time.
    Figure Legend Snippet: Proposed model for establishment of persistent MERS-CoV infection in bat cells. As with the stocks of most RNA viruses, the MERS-CoV inoculum is made up of the dominant W + virus as well as smaller numbers of variants, including variants with inactivating mutations in ORF5 (ΔORF5). The cells infected with the more cytolytic W + virus die, while the small number of cells infected with ΔORF5 MERS-CoV survive because of an ensuing antiviral response and the induction of anti-apoptotic processes. ΔORF5 MERS-CoV infected cells are resistant to infection with the W + virus and the W + virus is soon diluted out. After a process of cell death and recovery, ΔORF5 MERS-CoV infected cells survive and take over, leading to a culture of persistently infected cells that produce small, but consistent amounts of virus over time.

    Techniques Used: Infection

    MERS-CoV ΔORF5 mutant persistently infects bat (Efk) cells. ( a ) Schematic highlighting mutations (red arrows) in the MERS-CoV genome that were identified by sequencing the dominant virus strain in persistently infected bat (Efk) cells (passage 15) are shown. ( b ) Levels of W+ and ΔORF5 MERS-CoV replication in bat (Efk) and human (MRC5) cells. Expression levels of MERS-CoV upE gene (−(ΔCT gene − ΔCT GAPDH )), normalized to mock infected cells are shown (n = 4; Mean ± SD). *P = 0.015 and ***P = 0.0003 (Holm-Sidak t test with α = 0.05).
    Figure Legend Snippet: MERS-CoV ΔORF5 mutant persistently infects bat (Efk) cells. ( a ) Schematic highlighting mutations (red arrows) in the MERS-CoV genome that were identified by sequencing the dominant virus strain in persistently infected bat (Efk) cells (passage 15) are shown. ( b ) Levels of W+ and ΔORF5 MERS-CoV replication in bat (Efk) and human (MRC5) cells. Expression levels of MERS-CoV upE gene (−(ΔCT gene − ΔCT GAPDH )), normalized to mock infected cells are shown (n = 4; Mean ± SD). *P = 0.015 and ***P = 0.0003 (Holm-Sidak t test with α = 0.05).

    Techniques Used: Mutagenesis, Sequencing, Infection, Expressing

    Persistently infected bat cells are resistant to super-infection with wildtype or ΔORF5 MERS-CoV. ( a–c ) Efk cells persistently infected with MERS-CoV were superinfected with W + MERS-CoV (blue) and transcript levels for ( a ) upE, ( b ) ORF5 or ( c ) E. fuscus dipeptidyl peptidase 4 (DPP4) were measured. The expression levels of upE, ORF5 and DPP4 transcripts (−(ΔCT gene − ΔCT GAPDH )) were also measured in Efk cells that were persistently infected with MERS-CoV in the absence of additional virus (red) and naïve Efk cells infected with W + MERS-CoV (green), with respect to time 0 for input W + virus (n = 3; Mean ± SD). DPP4 qRT-PCR amplicons were analyzed on an agarose gel (gel inset) and a ratio of basal DPP4 transcript levels (−(ΔCT DPP4 − ΔCT GAPDH )) in naïve, uninfected Efk and MRC5 cells is shown (right panel). ***P
    Figure Legend Snippet: Persistently infected bat cells are resistant to super-infection with wildtype or ΔORF5 MERS-CoV. ( a–c ) Efk cells persistently infected with MERS-CoV were superinfected with W + MERS-CoV (blue) and transcript levels for ( a ) upE, ( b ) ORF5 or ( c ) E. fuscus dipeptidyl peptidase 4 (DPP4) were measured. The expression levels of upE, ORF5 and DPP4 transcripts (−(ΔCT gene − ΔCT GAPDH )) were also measured in Efk cells that were persistently infected with MERS-CoV in the absence of additional virus (red) and naïve Efk cells infected with W + MERS-CoV (green), with respect to time 0 for input W + virus (n = 3; Mean ± SD). DPP4 qRT-PCR amplicons were analyzed on an agarose gel (gel inset) and a ratio of basal DPP4 transcript levels (−(ΔCT DPP4 − ΔCT GAPDH )) in naïve, uninfected Efk and MRC5 cells is shown (right panel). ***P

    Techniques Used: Infection, Expressing, Quantitative RT-PCR, Agarose Gel Electrophoresis

    Bat cells can be persistently infected with MERS-CoV. ( a ) Big brown bat kidney cells (Efk) were infected with MERS-CoV (MOI = 0.01 TCID 50 /cell) for 12 days and then passaged weekly. Supernatant was collected during each passage to determine the presence of virus by titration on Vero cells, along with immunofluorescent and electron microscopic studies of infected cells. ( b ) Levels of MERS-CoV at different times following initial infection. ( c ) Phase contrast micrographs showing cytopathic effects on MERS-CoV infection and subsequent recovery of Efk cells at various time points. ( d ) Immunofluorescent images showing MERS-CoV nucleocapsid (N) protein in persistently infected Efk cells (bottom row; red arrows). The contrast for persistently infected Efk cells (inset) was adjusted to visualize low levels of protein. High MOI acute infection (middle row) and mock infection of Efk cells (top row) were used as positive and negative controls, respectively. Images were processed using ImageJ. ( e ) In-situ hybridization to detect the presence of MERS-CoV nucleoprotein RNA in persistently infected Efk cells. High, intermediate and low levels of MERS-CoV nucleoprotein RNA have been shown in the insets. Acutely infected (right) and mock infected (left) cells were used as positive and negative controls, respectively.
    Figure Legend Snippet: Bat cells can be persistently infected with MERS-CoV. ( a ) Big brown bat kidney cells (Efk) were infected with MERS-CoV (MOI = 0.01 TCID 50 /cell) for 12 days and then passaged weekly. Supernatant was collected during each passage to determine the presence of virus by titration on Vero cells, along with immunofluorescent and electron microscopic studies of infected cells. ( b ) Levels of MERS-CoV at different times following initial infection. ( c ) Phase contrast micrographs showing cytopathic effects on MERS-CoV infection and subsequent recovery of Efk cells at various time points. ( d ) Immunofluorescent images showing MERS-CoV nucleocapsid (N) protein in persistently infected Efk cells (bottom row; red arrows). The contrast for persistently infected Efk cells (inset) was adjusted to visualize low levels of protein. High MOI acute infection (middle row) and mock infection of Efk cells (top row) were used as positive and negative controls, respectively. Images were processed using ImageJ. ( e ) In-situ hybridization to detect the presence of MERS-CoV nucleoprotein RNA in persistently infected Efk cells. High, intermediate and low levels of MERS-CoV nucleoprotein RNA have been shown in the insets. Acutely infected (right) and mock infected (left) cells were used as positive and negative controls, respectively.

    Techniques Used: Infection, Titration, In Situ Hybridization

    IRF3 and MAP kinase-mediated signaling regulate persistent infection in Efk cells. Persistently infected Efk cells were transfected with siRNA targeting IRF3 mRNA and the subsequent effect on virus replication was measured. ( a) MERS-CoV titres in persistently infected bat cells 24 hours post treatment with IRF3-siRNA (red bar; n = 4; Mean ± SD). Scrambled siRNA (blue bar; control-siRNA) was used as a negative control. **P = 0.0049 (Unpaired t test with α = 0.05). ( b) Western blot for IRF3 and GAPDH in Efk cells treated or mock treated with IRF3 siRNA. (c) MERS-CoV upE transcript levels in Efk cells after treatment or mock treatment with MAPK inhibitor, URMC-99 for 24 and 48 hours. * P = 0.0053 ( Holm-Sidak t test with α=0.05). ( d) RERG transcript levels 48-hours post treatment with URMC-99 (n = 4; Mean ± SD). * P = 0.017 ( Holm-Sidak t test with α = 0.05). For full size gel images in (b) , see supplementary Fig. S3 .
    Figure Legend Snippet: IRF3 and MAP kinase-mediated signaling regulate persistent infection in Efk cells. Persistently infected Efk cells were transfected with siRNA targeting IRF3 mRNA and the subsequent effect on virus replication was measured. ( a) MERS-CoV titres in persistently infected bat cells 24 hours post treatment with IRF3-siRNA (red bar; n = 4; Mean ± SD). Scrambled siRNA (blue bar; control-siRNA) was used as a negative control. **P = 0.0049 (Unpaired t test with α = 0.05). ( b) Western blot for IRF3 and GAPDH in Efk cells treated or mock treated with IRF3 siRNA. (c) MERS-CoV upE transcript levels in Efk cells after treatment or mock treatment with MAPK inhibitor, URMC-99 for 24 and 48 hours. * P = 0.0053 ( Holm-Sidak t test with α=0.05). ( d) RERG transcript levels 48-hours post treatment with URMC-99 (n = 4; Mean ± SD). * P = 0.017 ( Holm-Sidak t test with α = 0.05). For full size gel images in (b) , see supplementary Fig. S3 .

    Techniques Used: Infection, Transfection, Negative Control, Western Blot

    2) Product Images from "Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model"

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model

    Journal: Vaccines

    doi: 10.3390/vaccines8040634

    Kinetics of serum and egg yolk anti-MERS COV-S IgY antibodies response of chickens after immunization with MERS COV-S recombinant protein compared with the adjuvant-immunized chicken (adjuvant control). Each week is represented by a pool of egg yolks of individual chicken in each group (S1-immunized and adjuvant-immunized).
    Figure Legend Snippet: Kinetics of serum and egg yolk anti-MERS COV-S IgY antibodies response of chickens after immunization with MERS COV-S recombinant protein compared with the adjuvant-immunized chicken (adjuvant control). Each week is represented by a pool of egg yolks of individual chicken in each group (S1-immunized and adjuvant-immunized).

    Techniques Used: Recombinant

    Recognition by anti-S1 IgY antibodies of viral antigen expressed in MERS-CoV-infected Vero E6 cells, using indirect immunofluorescence assay. ( A ) Vero E6 cells inoculated with MERS-CoV and stained with anti-S1 IgY antibodies and FITC-conjugated anti-chicken antibodies; and ( B ) control adjuvant IgY (Bright-field).
    Figure Legend Snippet: Recognition by anti-S1 IgY antibodies of viral antigen expressed in MERS-CoV-infected Vero E6 cells, using indirect immunofluorescence assay. ( A ) Vero E6 cells inoculated with MERS-CoV and stained with anti-S1 IgY antibodies and FITC-conjugated anti-chicken antibodies; and ( B ) control adjuvant IgY (Bright-field).

    Techniques Used: Infection, Immunofluorescence, Staining

    Dot blotting analysis. Purified anti-S1 IgY antibodies showed reactivity with different concentrations of the spike protein (S), S1, and receptor binding domain (RBD), but had no reactivity with nucleocapsid (NP) protein of MERS CoV.
    Figure Legend Snippet: Dot blotting analysis. Purified anti-S1 IgY antibodies showed reactivity with different concentrations of the spike protein (S), S1, and receptor binding domain (RBD), but had no reactivity with nucleocapsid (NP) protein of MERS CoV.

    Techniques Used: Purification, Binding Assay

    Examples of different concentrations of anti-S1 IgY antibodies tested against MERS-CoV on Vero-E6 cells examined by CPE. The IC100 neutralization of the antibody were determined as the reciprocal of the highest dilution at which no CPE was observed.
    Figure Legend Snippet: Examples of different concentrations of anti-S1 IgY antibodies tested against MERS-CoV on Vero-E6 cells examined by CPE. The IC100 neutralization of the antibody were determined as the reciprocal of the highest dilution at which no CPE was observed.

    Techniques Used: Neutralization

    ( A ) Viral titer in the lungs of MERS-CoV mice treated with anti-SI IgY antibodies and control IgY (adjuvant). ( B ) Body weight changes after MERS-CoV infection between anti-SI IgY antibodies and IgY of adjuvant control group. ( C – F ) Histopathology of the lungs from human dipeptidyl peptidase 4 (hDPP4)-transgenic mice on day 8 after inoculation with MERS-CoV. Representative histopathological findings of mice with the highest cellular infiltration in alveoli by H E staining ( C ) Massive mononuclear cell infiltrations including macrophages and lymphocytes with regenerated type II pneumocytes were seen in adjuvant control group (right column), but less in the anti-S1 IgY treated group (left column). Scale bars: 200 μm (upper row) and 20 μm (lower row). Al, alveoli; Br, bronchi; V, vessel. Detection of viral antigen in lung tissues of mice by immunohistochemistry ( D ) A few antigen positive cells were seen in the lungs of anti-S1 IgY treated group compared to adjuvant control group. Quantification of inflammation areas ( E ) The area of pulmonary lesion was determined based on the mean percentage of affected areas in each section of the collected lobes form each animal ( n = 8 or 6). Circles indicate averages from three observation lobes in each mouse. p = 0.1709 by Mann-Whitney test. Numbers of viral antigen positive cells in the alveoli ( F ) Data were obtained from 8 or 6 mice. Circles indicate averages of 5 observation fields in each mouse. * p = 0.0196 by Mann-Whitney test.
    Figure Legend Snippet: ( A ) Viral titer in the lungs of MERS-CoV mice treated with anti-SI IgY antibodies and control IgY (adjuvant). ( B ) Body weight changes after MERS-CoV infection between anti-SI IgY antibodies and IgY of adjuvant control group. ( C – F ) Histopathology of the lungs from human dipeptidyl peptidase 4 (hDPP4)-transgenic mice on day 8 after inoculation with MERS-CoV. Representative histopathological findings of mice with the highest cellular infiltration in alveoli by H E staining ( C ) Massive mononuclear cell infiltrations including macrophages and lymphocytes with regenerated type II pneumocytes were seen in adjuvant control group (right column), but less in the anti-S1 IgY treated group (left column). Scale bars: 200 μm (upper row) and 20 μm (lower row). Al, alveoli; Br, bronchi; V, vessel. Detection of viral antigen in lung tissues of mice by immunohistochemistry ( D ) A few antigen positive cells were seen in the lungs of anti-S1 IgY treated group compared to adjuvant control group. Quantification of inflammation areas ( E ) The area of pulmonary lesion was determined based on the mean percentage of affected areas in each section of the collected lobes form each animal ( n = 8 or 6). Circles indicate averages from three observation lobes in each mouse. p = 0.1709 by Mann-Whitney test. Numbers of viral antigen positive cells in the alveoli ( F ) Data were obtained from 8 or 6 mice. Circles indicate averages of 5 observation fields in each mouse. * p = 0.0196 by Mann-Whitney test.

    Techniques Used: Mouse Assay, Infection, Histopathology, Transgenic Assay, Staining, Immunohistochemistry, MANN-WHITNEY

    Western blot analysis of anti-MERS-COV rS1 IgY antibodies. (Left) The S1 protein of MERS-COV was subjected to SDS-PAGE under reducing conditions; (Right) Western blot analysis of the anti-S1 IgY antibody response. SDS gels were electrically transferred onto nitrocellulose membranes and probed with IgY from immunized and nonimmunized hens (marker: molecular maker; lane A: S1-immunized IgY; lane B: adjuvant-immunized IgY). The strips were processed separately and pasted beside each other for documentation.
    Figure Legend Snippet: Western blot analysis of anti-MERS-COV rS1 IgY antibodies. (Left) The S1 protein of MERS-COV was subjected to SDS-PAGE under reducing conditions; (Right) Western blot analysis of the anti-S1 IgY antibody response. SDS gels were electrically transferred onto nitrocellulose membranes and probed with IgY from immunized and nonimmunized hens (marker: molecular maker; lane A: S1-immunized IgY; lane B: adjuvant-immunized IgY). The strips were processed separately and pasted beside each other for documentation.

    Techniques Used: Western Blot, SDS Page, Marker

    Evaluation of the neutralizing potential of anti-S1 IgY antibodies, using plaque reduction neutralization test. ( A ) MERS-CoV (MOI 0.01) was incubated with different concentrations of anti-S1 IgY antibodies and added to Vero E6 cells. After virus adsorption, agar medium was added to the Vero E6 cells, and the plaques that formed were stained with crystal violet, each IgY concentration was tested in triplicate. ( B ) Percent inhibition of anti-S1 IgY antibodies with different concentrations. The best fit equation is:
    Figure Legend Snippet: Evaluation of the neutralizing potential of anti-S1 IgY antibodies, using plaque reduction neutralization test. ( A ) MERS-CoV (MOI 0.01) was incubated with different concentrations of anti-S1 IgY antibodies and added to Vero E6 cells. After virus adsorption, agar medium was added to the Vero E6 cells, and the plaques that formed were stained with crystal violet, each IgY concentration was tested in triplicate. ( B ) Percent inhibition of anti-S1 IgY antibodies with different concentrations. The best fit equation is:

    Techniques Used: Plaque Reduction Neutralization Test, Incubation, Adsorption, Staining, Concentration Assay, Inhibition

    3) Product Images from "Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus"

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-64264-1

    ΔORF5 MERS-CoV infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P
    Figure Legend Snippet: ΔORF5 MERS-CoV infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P

    Techniques Used: Infection

    MERS-CoV gene expression varies between acute and persistently infected bat cells. RNA from persistently infected Efk cells and acutely infected Efk cells were harvested at several time points and MERS-CoV genome quantities (upE levels) and gene expression levels were analyzed by real time quantitative PCR. ( a–i) Genome and gene expression (−(ΔCT gene − ΔCT GAPDH )) levels for MERS-CoV upE, S, ORF3, ORF4a, ORF4b, ORF5, E, M and N genes in acute (green) vs. persistent (purple) infections at 0, 12, 24 and 48 hours post-infection or seeding, respectively (n = 4; Mean ± SD). ***P
    Figure Legend Snippet: MERS-CoV gene expression varies between acute and persistently infected bat cells. RNA from persistently infected Efk cells and acutely infected Efk cells were harvested at several time points and MERS-CoV genome quantities (upE levels) and gene expression levels were analyzed by real time quantitative PCR. ( a–i) Genome and gene expression (−(ΔCT gene − ΔCT GAPDH )) levels for MERS-CoV upE, S, ORF3, ORF4a, ORF4b, ORF5, E, M and N genes in acute (green) vs. persistent (purple) infections at 0, 12, 24 and 48 hours post-infection or seeding, respectively (n = 4; Mean ± SD). ***P

    Techniques Used: Expressing, Infection, Real-time Polymerase Chain Reaction

    Proposed model for establishment of persistent MERS-CoV infection in bat cells. As with the stocks of most RNA viruses, the MERS-CoV inoculum is made up of the dominant W + virus as well as smaller numbers of variants, including variants with inactivating mutations in ORF5 (ΔORF5). The cells infected with the more cytolytic W + virus die, while the small number of cells infected with ΔORF5 MERS-CoV survive because of an ensuing antiviral response and the induction of anti-apoptotic processes. ΔORF5 MERS-CoV infected cells are resistant to infection with the W + virus and the W + virus is soon diluted out. After a process of cell death and recovery, ΔORF5 MERS-CoV infected cells survive and take over, leading to a culture of persistently infected cells that produce small, but consistent amounts of virus over time.
    Figure Legend Snippet: Proposed model for establishment of persistent MERS-CoV infection in bat cells. As with the stocks of most RNA viruses, the MERS-CoV inoculum is made up of the dominant W + virus as well as smaller numbers of variants, including variants with inactivating mutations in ORF5 (ΔORF5). The cells infected with the more cytolytic W + virus die, while the small number of cells infected with ΔORF5 MERS-CoV survive because of an ensuing antiviral response and the induction of anti-apoptotic processes. ΔORF5 MERS-CoV infected cells are resistant to infection with the W + virus and the W + virus is soon diluted out. After a process of cell death and recovery, ΔORF5 MERS-CoV infected cells survive and take over, leading to a culture of persistently infected cells that produce small, but consistent amounts of virus over time.

    Techniques Used: Infection

    MERS-CoV ΔORF5 mutant persistently infects bat (Efk) cells. ( a ) Schematic highlighting mutations (red arrows) in the MERS-CoV genome that were identified by sequencing the dominant virus strain in persistently infected bat (Efk) cells (passage 15) are shown. ( b ) Levels of W+ and ΔORF5 MERS-CoV replication in bat (Efk) and human (MRC5) cells. Expression levels of MERS-CoV upE gene (−(ΔCT gene − ΔCT GAPDH )), normalized to mock infected cells are shown (n = 4; Mean ± SD). *P = 0.015 and ***P = 0.0003 (Holm-Sidak t test with α = 0.05).
    Figure Legend Snippet: MERS-CoV ΔORF5 mutant persistently infects bat (Efk) cells. ( a ) Schematic highlighting mutations (red arrows) in the MERS-CoV genome that were identified by sequencing the dominant virus strain in persistently infected bat (Efk) cells (passage 15) are shown. ( b ) Levels of W+ and ΔORF5 MERS-CoV replication in bat (Efk) and human (MRC5) cells. Expression levels of MERS-CoV upE gene (−(ΔCT gene − ΔCT GAPDH )), normalized to mock infected cells are shown (n = 4; Mean ± SD). *P = 0.015 and ***P = 0.0003 (Holm-Sidak t test with α = 0.05).

    Techniques Used: Mutagenesis, Sequencing, Infection, Expressing

    Persistently infected bat cells are resistant to super-infection with wildtype or ΔORF5 MERS-CoV. ( a–c ) Efk cells persistently infected with MERS-CoV were superinfected with W + MERS-CoV (blue) and transcript levels for ( a ) upE, ( b ) ORF5 or ( c ) E. fuscus dipeptidyl peptidase 4 (DPP4) were measured. The expression levels of upE, ORF5 and DPP4 transcripts (−(ΔCT gene − ΔCT GAPDH )) were also measured in Efk cells that were persistently infected with MERS-CoV in the absence of additional virus (red) and naïve Efk cells infected with W + MERS-CoV (green), with respect to time 0 for input W + virus (n = 3; Mean ± SD). DPP4 qRT-PCR amplicons were analyzed on an agarose gel (gel inset) and a ratio of basal DPP4 transcript levels (−(ΔCT DPP4 − ΔCT GAPDH )) in naïve, uninfected Efk and MRC5 cells is shown (right panel). ***P
    Figure Legend Snippet: Persistently infected bat cells are resistant to super-infection with wildtype or ΔORF5 MERS-CoV. ( a–c ) Efk cells persistently infected with MERS-CoV were superinfected with W + MERS-CoV (blue) and transcript levels for ( a ) upE, ( b ) ORF5 or ( c ) E. fuscus dipeptidyl peptidase 4 (DPP4) were measured. The expression levels of upE, ORF5 and DPP4 transcripts (−(ΔCT gene − ΔCT GAPDH )) were also measured in Efk cells that were persistently infected with MERS-CoV in the absence of additional virus (red) and naïve Efk cells infected with W + MERS-CoV (green), with respect to time 0 for input W + virus (n = 3; Mean ± SD). DPP4 qRT-PCR amplicons were analyzed on an agarose gel (gel inset) and a ratio of basal DPP4 transcript levels (−(ΔCT DPP4 − ΔCT GAPDH )) in naïve, uninfected Efk and MRC5 cells is shown (right panel). ***P

    Techniques Used: Infection, Expressing, Quantitative RT-PCR, Agarose Gel Electrophoresis

    Bat cells can be persistently infected with MERS-CoV. ( a ) Big brown bat kidney cells (Efk) were infected with MERS-CoV (MOI = 0.01 TCID 50 /cell) for 12 days and then passaged weekly. Supernatant was collected during each passage to determine the presence of virus by titration on Vero cells, along with immunofluorescent and electron microscopic studies of infected cells. ( b ) Levels of MERS-CoV at different times following initial infection. ( c ) Phase contrast micrographs showing cytopathic effects on MERS-CoV infection and subsequent recovery of Efk cells at various time points. ( d ) Immunofluorescent images showing MERS-CoV nucleocapsid (N) protein in persistently infected Efk cells (bottom row; red arrows). The contrast for persistently infected Efk cells (inset) was adjusted to visualize low levels of protein. High MOI acute infection (middle row) and mock infection of Efk cells (top row) were used as positive and negative controls, respectively. Images were processed using ImageJ. ( e ) In-situ hybridization to detect the presence of MERS-CoV nucleoprotein RNA in persistently infected Efk cells. High, intermediate and low levels of MERS-CoV nucleoprotein RNA have been shown in the insets. Acutely infected (right) and mock infected (left) cells were used as positive and negative controls, respectively.
    Figure Legend Snippet: Bat cells can be persistently infected with MERS-CoV. ( a ) Big brown bat kidney cells (Efk) were infected with MERS-CoV (MOI = 0.01 TCID 50 /cell) for 12 days and then passaged weekly. Supernatant was collected during each passage to determine the presence of virus by titration on Vero cells, along with immunofluorescent and electron microscopic studies of infected cells. ( b ) Levels of MERS-CoV at different times following initial infection. ( c ) Phase contrast micrographs showing cytopathic effects on MERS-CoV infection and subsequent recovery of Efk cells at various time points. ( d ) Immunofluorescent images showing MERS-CoV nucleocapsid (N) protein in persistently infected Efk cells (bottom row; red arrows). The contrast for persistently infected Efk cells (inset) was adjusted to visualize low levels of protein. High MOI acute infection (middle row) and mock infection of Efk cells (top row) were used as positive and negative controls, respectively. Images were processed using ImageJ. ( e ) In-situ hybridization to detect the presence of MERS-CoV nucleoprotein RNA in persistently infected Efk cells. High, intermediate and low levels of MERS-CoV nucleoprotein RNA have been shown in the insets. Acutely infected (right) and mock infected (left) cells were used as positive and negative controls, respectively.

    Techniques Used: Infection, Titration, In Situ Hybridization

    IRF3 and MAP kinase-mediated signaling regulate persistent infection in Efk cells. Persistently infected Efk cells were transfected with siRNA targeting IRF3 mRNA and the subsequent effect on virus replication was measured. ( a) MERS-CoV titres in persistently infected bat cells 24 hours post treatment with IRF3-siRNA (red bar; n = 4; Mean ± SD). Scrambled siRNA (blue bar; control-siRNA) was used as a negative control. **P = 0.0049 (Unpaired t test with α = 0.05). ( b) Western blot for IRF3 and GAPDH in Efk cells treated or mock treated with IRF3 siRNA. (c) MERS-CoV upE transcript levels in Efk cells after treatment or mock treatment with MAPK inhibitor, URMC-99 for 24 and 48 hours. * P = 0.0053 ( Holm-Sidak t test with α=0.05). ( d) RERG transcript levels 48-hours post treatment with URMC-99 (n = 4; Mean ± SD). * P = 0.017 ( Holm-Sidak t test with α = 0.05). For full size gel images in (b) , see supplementary Fig. S3 .
    Figure Legend Snippet: IRF3 and MAP kinase-mediated signaling regulate persistent infection in Efk cells. Persistently infected Efk cells were transfected with siRNA targeting IRF3 mRNA and the subsequent effect on virus replication was measured. ( a) MERS-CoV titres in persistently infected bat cells 24 hours post treatment with IRF3-siRNA (red bar; n = 4; Mean ± SD). Scrambled siRNA (blue bar; control-siRNA) was used as a negative control. **P = 0.0049 (Unpaired t test with α = 0.05). ( b) Western blot for IRF3 and GAPDH in Efk cells treated or mock treated with IRF3 siRNA. (c) MERS-CoV upE transcript levels in Efk cells after treatment or mock treatment with MAPK inhibitor, URMC-99 for 24 and 48 hours. * P = 0.0053 ( Holm-Sidak t test with α=0.05). ( d) RERG transcript levels 48-hours post treatment with URMC-99 (n = 4; Mean ± SD). * P = 0.017 ( Holm-Sidak t test with α = 0.05). For full size gel images in (b) , see supplementary Fig. S3 .

    Techniques Used: Infection, Transfection, Negative Control, Western Blot

    4) Product Images from "Immunotherapeutic Efficacy of IgY Antibodies Targeting the Full-Length Spike Protein in an Animal Model of Middle East Respiratory Syndrome Coronavirus Infection"

    Article Title: Immunotherapeutic Efficacy of IgY Antibodies Targeting the Full-Length Spike Protein in an Animal Model of Middle East Respiratory Syndrome Coronavirus Infection

    Journal: Pharmaceuticals

    doi: 10.3390/ph14060511

    ( A ) SDS-PAGE profile of anti-MERS-CoV S IgY antibodies. The two IgY chains appeared using 10% resolving SDS-PAGE gel. The molecular weight of the heavy chain is 68 kDa, and the molecular weight of the light chain is 27 kDa. ( B ) Western blot identification of IgY using HRP-conjugated rabbit anti-chicken IgY heavy and light. Remaining bands might represent other antibodies or protein fragments of unknown origin.
    Figure Legend Snippet: ( A ) SDS-PAGE profile of anti-MERS-CoV S IgY antibodies. The two IgY chains appeared using 10% resolving SDS-PAGE gel. The molecular weight of the heavy chain is 68 kDa, and the molecular weight of the light chain is 27 kDa. ( B ) Western blot identification of IgY using HRP-conjugated rabbit anti-chicken IgY heavy and light. Remaining bands might represent other antibodies or protein fragments of unknown origin.

    Techniques Used: SDS Page, Molecular Weight, Western Blot

    Dot blotting analysis. Purified anti-S IgY antibodies showed reactivity with different concentrations of the S, S1, and receptor-binding domain proteins but had no reactivity with the nucleocapsid protein of MERS-CoV.
    Figure Legend Snippet: Dot blotting analysis. Purified anti-S IgY antibodies showed reactivity with different concentrations of the S, S1, and receptor-binding domain proteins but had no reactivity with the nucleocapsid protein of MERS-CoV.

    Techniques Used: Purification, Binding Assay

    Western blot analysis of anti-MERS-CoV S IgY antibodies. ( A ) The S protein of MERS-CoV subjected to SDS-PAGE under reducing conditions. ( B ) Western blot analysis of the anti-S IgY antibody response.
    Figure Legend Snippet: Western blot analysis of anti-MERS-CoV S IgY antibodies. ( A ) The S protein of MERS-CoV subjected to SDS-PAGE under reducing conditions. ( B ) Western blot analysis of the anti-S IgY antibody response.

    Techniques Used: Western Blot, SDS Page

    Cytopathic effect of different concentrations of anti-S IgY antibodies against MERS-CoV in Vero-E6 cells. The IC 100 neutralization of the antibody was determined as the reciprocal of the highest dilution at which no cytopathic effect was observed.
    Figure Legend Snippet: Cytopathic effect of different concentrations of anti-S IgY antibodies against MERS-CoV in Vero-E6 cells. The IC 100 neutralization of the antibody was determined as the reciprocal of the highest dilution at which no cytopathic effect was observed.

    Techniques Used: Neutralization

    ( A ) Viral titers in lung homogenates of MERS-CoV infected mice at 1-, 3-, and 5-days after inoculation ( n = 4 per group). Mice were treated with anti-S IgY antibodies or adjuvant only. The detection limit was 10^1.5 TCID 50 /g of tissue. ( B ) Body weight changes between mice with anti-S IgY antibodies and the adjuvant-only controls after MERS-CoV infection. ( C – F ) Histopathology of the lungs from human dipeptidyl peptidase 4-transgenic mice on day 8 after infection with MERS-CoV. ( C ) Representative histopathological findings of mice with highest cellular infiltration in the alveoli, identified using hematoxylin and eosin staining. Massive mononuclear cell infiltrations, including macrophages and lymphocytes with regenerated type II pneumocytes, were observed in the control group (right column) but slightly less in the group treated with anti-S IgY (left column). Scale bars: 200 μm (upper row) and 20 μm (lower row). Al, alveoli; Br, bronchi; V, vessel. ( D ) Quantification of inflammation areas. Pulmonary lesion areas were determined based on the mean percentage of affected areas in each section of the collected lobes from six animals. Circles indicate averages from three observation lobes in each mouse ( p = 0.041 by Mann-Whitney test). ( E ) Detection of viral antigen in the lung tissues of mice, determined by immunohistochemistry. Antigen-positive cells were observed less frequently in the lungs of the group treated with anti-S-IgY, compared to the adjuvant-only controls. Scale bars: 20 μm. ( F ) Numbers of viral-antigen-positive cells in the alveoli from six mice. Circles indicate averages of five observation fields in each mouse ( p = 0.258 by Mann-Whitney test). The asterisk indicates statistical significance.
    Figure Legend Snippet: ( A ) Viral titers in lung homogenates of MERS-CoV infected mice at 1-, 3-, and 5-days after inoculation ( n = 4 per group). Mice were treated with anti-S IgY antibodies or adjuvant only. The detection limit was 10^1.5 TCID 50 /g of tissue. ( B ) Body weight changes between mice with anti-S IgY antibodies and the adjuvant-only controls after MERS-CoV infection. ( C – F ) Histopathology of the lungs from human dipeptidyl peptidase 4-transgenic mice on day 8 after infection with MERS-CoV. ( C ) Representative histopathological findings of mice with highest cellular infiltration in the alveoli, identified using hematoxylin and eosin staining. Massive mononuclear cell infiltrations, including macrophages and lymphocytes with regenerated type II pneumocytes, were observed in the control group (right column) but slightly less in the group treated with anti-S IgY (left column). Scale bars: 200 μm (upper row) and 20 μm (lower row). Al, alveoli; Br, bronchi; V, vessel. ( D ) Quantification of inflammation areas. Pulmonary lesion areas were determined based on the mean percentage of affected areas in each section of the collected lobes from six animals. Circles indicate averages from three observation lobes in each mouse ( p = 0.041 by Mann-Whitney test). ( E ) Detection of viral antigen in the lung tissues of mice, determined by immunohistochemistry. Antigen-positive cells were observed less frequently in the lungs of the group treated with anti-S-IgY, compared to the adjuvant-only controls. Scale bars: 20 μm. ( F ) Numbers of viral-antigen-positive cells in the alveoli from six mice. Circles indicate averages of five observation fields in each mouse ( p = 0.258 by Mann-Whitney test). The asterisk indicates statistical significance.

    Techniques Used: Infection, Mouse Assay, Histopathology, Transgenic Assay, Staining, MANN-WHITNEY, Immunohistochemistry

    Kinetics of chicken serum and egg yolk antibody response to anti-MERS-CoV S IgY after infection with MERS-CoV S recombinant protein, compared with adjuvant-only controls. Each week is represented by a pool of egg yolks of individual chickens from each group (S-immunized and adjuvant-only).
    Figure Legend Snippet: Kinetics of chicken serum and egg yolk antibody response to anti-MERS-CoV S IgY after infection with MERS-CoV S recombinant protein, compared with adjuvant-only controls. Each week is represented by a pool of egg yolks of individual chickens from each group (S-immunized and adjuvant-only).

    Techniques Used: Infection, Recombinant

    Related Articles

    Immunohistochemistry:

    Article Title: Acute Respiratory Infection in Human Dipeptidyl Peptidase 4-Transgenic Mice Infected with Middle East Respiratory Syndrome Coronavirus
    Article Snippet: .. For IHC, antigen retrieval of formalin-fixed mouse tissue sections was performed by autoclaving at 121°C for 10 min in retrieval solution at pH 6.0 (Nichirei, Tokyo, Japan). hDPP4 and MERS-CoV antigens were detected using a standard immunoperoxidase method and a goat anti-hDPP4 antibody (R & D Systems) and a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological, Inc., Beijing, China). ..

    Article Title: The characteristics of hDPP4 transgenic mice subjected to aerosol MERS coronavirus infection via an animal nose‐only exposure device. The characteristics of hDPP4 transgenic mice subjected to aerosol MERS coronavirus infection via an animal nose‐only exposure device
    Article Snippet: .. Immunohistochemical staining was performed to assess the expression of a viral antigen using a rabbit two‐step detection kit (Zhongshan Golden Bridge Biotechnology Co., Ltd) with a rabbit polyclonal anti‐MERS‐CoV nucleoprotein (NP) antibody (Sino Biological Inc). ..

    Staining:

    Article Title: The characteristics of hDPP4 transgenic mice subjected to aerosol MERS coronavirus infection via an animal nose‐only exposure device. The characteristics of hDPP4 transgenic mice subjected to aerosol MERS coronavirus infection via an animal nose‐only exposure device
    Article Snippet: .. Immunohistochemical staining was performed to assess the expression of a viral antigen using a rabbit two‐step detection kit (Zhongshan Golden Bridge Biotechnology Co., Ltd) with a rabbit polyclonal anti‐MERS‐CoV nucleoprotein (NP) antibody (Sino Biological Inc). ..

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus
    Article Snippet: .. Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302). ..

    Article Title: Anti-S1 MERS-COV IgY Specific Antibodies Decreases Lung Inflammation and Viral Antigen Positive Cells in the Human Transgenic Mouse Model
    Article Snippet: .. MERS-CoV antigens were detected utilizing a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R); Nichirei) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01; Sino Biological Inc., Beijing, China). ..

    Article Title: Immunotherapeutic Efficacy of IgY Antibodies Targeting the Full-Length Spike Protein in an Animal Model of Middle East Respiratory Syndrome Coronavirus Infection
    Article Snippet: .. MERS-CoV antigens were detected using a polymer-based detection system (Nichirei-Histofine Simple Stain Mouse MAX PO(R), Nichirei, Tokyo, Japan) with a rabbit anti-MERS-CoV nucleocapsid antibody (40068-RP01, Sino Biological Inc., Beijing, China). ..

    Expressing:

    Article Title: The characteristics of hDPP4 transgenic mice subjected to aerosol MERS coronavirus infection via an animal nose‐only exposure device. The characteristics of hDPP4 transgenic mice subjected to aerosol MERS coronavirus infection via an animal nose‐only exposure device
    Article Snippet: .. Immunohistochemical staining was performed to assess the expression of a viral antigen using a rabbit two‐step detection kit (Zhongshan Golden Bridge Biotechnology Co., Ltd) with a rabbit polyclonal anti‐MERS‐CoV nucleoprotein (NP) antibody (Sino Biological Inc). ..

    other:

    Article Title: Middle East respiratory syndrome coronavirus shows poor replication but significant induction of antiviral responses in human monocyte-derived macrophages and dendritic cells
    Article Snippet: Commercial rabbit antibodies against MERS-CoV N protein (1 : 1000 dilution; Sino Biological), human DPP4 (1 : 1000 dilution; AbCam) and actin (1 : 500 dilution; Santa Cruz Biotechnology) were used according to the manufacturer's instructions.

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    Sino Biological polyclonal rabbit anti mers cov nucleocapsid antibodies
    Representative HE findings in respiratory epithelium of the nasal turbinates of <t>MERS-CoV-infected</t> alpacas and immunohistochemical reactions of the commercially available positive controls for comparison. (A) Multifocal infiltration of lamina propria and submucosa by moderate numbers of lymphocytes, macrophages, and single neutrophilic granulocytes (asterisk). (B) Exocytosis of single neutrophilic granulocytes (grey arrow). (C, D) Abundant viral antigen was detected multifocally in the cytoplasm and along the apical membranous region of epithelial cells using a monoclonal mouse (C, Sino Biological Inc.) and <t>polyclonal</t> rabbit anti-MERS-CoV nucleocapsid antibody (D, Sino Biological Inc.) with citrate pretreatment, in a dilution of 1:70 and 1:2,000, respectively. Both antibodies exhibited a similar staining intensity. (A, B) HE staining; 400x, (C, D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.
    Polyclonal Rabbit Anti Mers Cov Nucleocapsid Antibodies, 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/polyclonal rabbit anti mers cov nucleocapsid antibodies/product/Sino Biological
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    polyclonal rabbit anti mers cov nucleocapsid antibodies - by Bioz Stars, 2021-09
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    94
    Sino Biological rabbit polyclonal nucleoprotein antibody
    Immunohistochemical labelling of FFPE SARS-CoV and SARS-CoV-2 infected cells and uninfected cells. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody (a–c) , SARS-CoV nucleoprotein rabbit <t>polyclonal</t> antibody (d–f) and double-stranded RNA (dsRNA) rabbit monoclonal antibody (g–i) . Scale bars, 20 µm.
    Rabbit Polyclonal Nucleoprotein 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
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    94
    Sino Biological mers cov nucleocapsid protein antibody rabbit pab
    IHC detection of virus antigen expression in mouse tissue after challenge with <t>MERS-CoV.</t> Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.
    Mers Cov Nucleocapsid Protein Antibody Rabbit Pab, 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/mers cov nucleocapsid protein antibody rabbit pab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
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    mers cov nucleocapsid protein antibody rabbit pab - by Bioz Stars, 2021-09
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    Representative HE findings in respiratory epithelium of the nasal turbinates of MERS-CoV-infected alpacas and immunohistochemical reactions of the commercially available positive controls for comparison. (A) Multifocal infiltration of lamina propria and submucosa by moderate numbers of lymphocytes, macrophages, and single neutrophilic granulocytes (asterisk). (B) Exocytosis of single neutrophilic granulocytes (grey arrow). (C, D) Abundant viral antigen was detected multifocally in the cytoplasm and along the apical membranous region of epithelial cells using a monoclonal mouse (C, Sino Biological Inc.) and polyclonal rabbit anti-MERS-CoV nucleocapsid antibody (D, Sino Biological Inc.) with citrate pretreatment, in a dilution of 1:70 and 1:2,000, respectively. Both antibodies exhibited a similar staining intensity. (A, B) HE staining; 400x, (C, D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Journal: Veterinary Immunology and Immunopathology

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

    doi: 10.1016/j.vetimm.2019.109939

    Figure Lengend Snippet: Representative HE findings in respiratory epithelium of the nasal turbinates of MERS-CoV-infected alpacas and immunohistochemical reactions of the commercially available positive controls for comparison. (A) Multifocal infiltration of lamina propria and submucosa by moderate numbers of lymphocytes, macrophages, and single neutrophilic granulocytes (asterisk). (B) Exocytosis of single neutrophilic granulocytes (grey arrow). (C, D) Abundant viral antigen was detected multifocally in the cytoplasm and along the apical membranous region of epithelial cells using a monoclonal mouse (C, Sino Biological Inc.) and polyclonal rabbit anti-MERS-CoV nucleocapsid antibody (D, Sino Biological Inc.) with citrate pretreatment, in a dilution of 1:70 and 1:2,000, respectively. Both antibodies exhibited a similar staining intensity. (A, B) HE staining; 400x, (C, D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Article Snippet: For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively.

    Techniques: Infection, Immunohistochemistry, Staining, Avidin-Biotin Assay

    Representative positive immunohistochemical reactions for human monoclonal antibodies. (A) Antibody 1.2g5 with citrate pretreatment exhibited a strong multifocal MERS-CoV antigen specific signal in the cytoplasm of ciliated respiratory epithelial cells (arrow) and segmentally along the apical membranous region (arrow heads). (B) The same protocol without pretreatment revealed a much fainter but similarly distributed staining of cytoplasm (arrow) and ciliary base (arrow heads). (C) Antibody 1.10f3 with citrate pretreatment displayed a strong diffuse background staining and only few positively stained cilia (arrowhead). Single intraepithelial plasma cells displayed a strong false positive intracytoplasmic staining (white arrow). (D) Antibody 1.6c7 with citrate pretreatment exhibited also a strong diffuse background staining and only few positive staining cilia (arrow heads). (A–D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Journal: Veterinary Immunology and Immunopathology

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

    doi: 10.1016/j.vetimm.2019.109939

    Figure Lengend Snippet: Representative positive immunohistochemical reactions for human monoclonal antibodies. (A) Antibody 1.2g5 with citrate pretreatment exhibited a strong multifocal MERS-CoV antigen specific signal in the cytoplasm of ciliated respiratory epithelial cells (arrow) and segmentally along the apical membranous region (arrow heads). (B) The same protocol without pretreatment revealed a much fainter but similarly distributed staining of cytoplasm (arrow) and ciliary base (arrow heads). (C) Antibody 1.10f3 with citrate pretreatment displayed a strong diffuse background staining and only few positively stained cilia (arrowhead). Single intraepithelial plasma cells displayed a strong false positive intracytoplasmic staining (white arrow). (D) Antibody 1.6c7 with citrate pretreatment exhibited also a strong diffuse background staining and only few positive staining cilia (arrow heads). (A–D) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Article Snippet: For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively.

    Techniques: Immunohistochemistry, Staining, Avidin-Biotin Assay

    Representative negative immunohistochemical reactions for human monoclonal antibodies. (A–E) Specific staining for MERS-CoV antigen was absent for the antibodies 1.6f9 (A), 4.6e10 (B), 7.7g6 (C), 1.8e5 (D), and 3.5g6 (E), respectively. Reactions were accompanied by mild to severe non-specific, intracytoplasmic background staining. (A–E) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Journal: Veterinary Immunology and Immunopathology

    Article Title: Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein

    doi: 10.1016/j.vetimm.2019.109939

    Figure Lengend Snippet: Representative negative immunohistochemical reactions for human monoclonal antibodies. (A–E) Specific staining for MERS-CoV antigen was absent for the antibodies 1.6f9 (A), 4.6e10 (B), 7.7g6 (C), 1.8e5 (D), and 3.5g6 (E), respectively. Reactions were accompanied by mild to severe non-specific, intracytoplasmic background staining. (A–E) Avidin-biotin-peroxidase complex method with 3,3′-diaminobenzidine as chromogen; 400x.

    Article Snippet: For positive control and elucidation of virus distribution, additional sections were stained with two different commercially available, previously published monoclonal mouse and polyclonal rabbit anti-MERS-CoV nucleocapsid antibodies (Sino Biological Inc.; ; ), respectively.

    Techniques: Immunohistochemistry, Staining, Avidin-Biotin Assay

    Immunohistochemical labelling of FFPE SARS-CoV and SARS-CoV-2 infected cells and uninfected cells. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody (a–c) , SARS-CoV nucleoprotein rabbit polyclonal antibody (d–f) and double-stranded RNA (dsRNA) rabbit monoclonal antibody (g–i) . Scale bars, 20 µm.

    Journal: Scientific Reports

    Article Title: Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens

    doi: 10.1038/s41598-020-78949-0

    Figure Lengend Snippet: Immunohistochemical labelling of FFPE SARS-CoV and SARS-CoV-2 infected cells and uninfected cells. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody (a–c) , SARS-CoV nucleoprotein rabbit polyclonal antibody (d–f) and double-stranded RNA (dsRNA) rabbit monoclonal antibody (g–i) . Scale bars, 20 µm.

    Article Snippet: The rabbit monoclonal (mAb; Sino Biological 40150-R007) spike antibody is compatible with either pH9 or 6 retrieval method, whereas rabbit polyclonal nucleoprotein antibody (Sino Biological, 40,143-T62) can be applied on virus antigen retrieved using either heated pH6 buffer or proteinase (Table ).

    Techniques: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Infection, Immunodetection

    ΔORF5 MERS-CoV infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: ΔORF5 MERS-CoV infection induces higher levels of IFNβ and ISGs in bat cells. ( a–e ) Transcript levels of IFNβ and interferon stimulated genes (ISGs), IFI6, GBP1, Mx1 and MDA5 in Efk and MRC5 cells infected with W+ or ΔORF5 MERS-CoV. (f) DPP4 transcript levels in W+ or ΔORF5 MERS-CoV infected Efk cells (n = 4; Mean ± SD). Bars represent average fold changes (2 −ΔΔCT ) in transcript levels compared to mock infected cells and normalized to GAPDH levels in each sample (n = 4; Mean ± SD). *P

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Infection

    MERS-CoV gene expression varies between acute and persistently infected bat cells. RNA from persistently infected Efk cells and acutely infected Efk cells were harvested at several time points and MERS-CoV genome quantities (upE levels) and gene expression levels were analyzed by real time quantitative PCR. ( a–i) Genome and gene expression (−(ΔCT gene − ΔCT GAPDH )) levels for MERS-CoV upE, S, ORF3, ORF4a, ORF4b, ORF5, E, M and N genes in acute (green) vs. persistent (purple) infections at 0, 12, 24 and 48 hours post-infection or seeding, respectively (n = 4; Mean ± SD). ***P

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: MERS-CoV gene expression varies between acute and persistently infected bat cells. RNA from persistently infected Efk cells and acutely infected Efk cells were harvested at several time points and MERS-CoV genome quantities (upE levels) and gene expression levels were analyzed by real time quantitative PCR. ( a–i) Genome and gene expression (−(ΔCT gene − ΔCT GAPDH )) levels for MERS-CoV upE, S, ORF3, ORF4a, ORF4b, ORF5, E, M and N genes in acute (green) vs. persistent (purple) infections at 0, 12, 24 and 48 hours post-infection or seeding, respectively (n = 4; Mean ± SD). ***P

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Expressing, Infection, Real-time Polymerase Chain Reaction

    Proposed model for establishment of persistent MERS-CoV infection in bat cells. As with the stocks of most RNA viruses, the MERS-CoV inoculum is made up of the dominant W + virus as well as smaller numbers of variants, including variants with inactivating mutations in ORF5 (ΔORF5). The cells infected with the more cytolytic W + virus die, while the small number of cells infected with ΔORF5 MERS-CoV survive because of an ensuing antiviral response and the induction of anti-apoptotic processes. ΔORF5 MERS-CoV infected cells are resistant to infection with the W + virus and the W + virus is soon diluted out. After a process of cell death and recovery, ΔORF5 MERS-CoV infected cells survive and take over, leading to a culture of persistently infected cells that produce small, but consistent amounts of virus over time.

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: Proposed model for establishment of persistent MERS-CoV infection in bat cells. As with the stocks of most RNA viruses, the MERS-CoV inoculum is made up of the dominant W + virus as well as smaller numbers of variants, including variants with inactivating mutations in ORF5 (ΔORF5). The cells infected with the more cytolytic W + virus die, while the small number of cells infected with ΔORF5 MERS-CoV survive because of an ensuing antiviral response and the induction of anti-apoptotic processes. ΔORF5 MERS-CoV infected cells are resistant to infection with the W + virus and the W + virus is soon diluted out. After a process of cell death and recovery, ΔORF5 MERS-CoV infected cells survive and take over, leading to a culture of persistently infected cells that produce small, but consistent amounts of virus over time.

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Infection

    MERS-CoV ΔORF5 mutant persistently infects bat (Efk) cells. ( a ) Schematic highlighting mutations (red arrows) in the MERS-CoV genome that were identified by sequencing the dominant virus strain in persistently infected bat (Efk) cells (passage 15) are shown. ( b ) Levels of W+ and ΔORF5 MERS-CoV replication in bat (Efk) and human (MRC5) cells. Expression levels of MERS-CoV upE gene (−(ΔCT gene − ΔCT GAPDH )), normalized to mock infected cells are shown (n = 4; Mean ± SD). *P = 0.015 and ***P = 0.0003 (Holm-Sidak t test with α = 0.05).

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: MERS-CoV ΔORF5 mutant persistently infects bat (Efk) cells. ( a ) Schematic highlighting mutations (red arrows) in the MERS-CoV genome that were identified by sequencing the dominant virus strain in persistently infected bat (Efk) cells (passage 15) are shown. ( b ) Levels of W+ and ΔORF5 MERS-CoV replication in bat (Efk) and human (MRC5) cells. Expression levels of MERS-CoV upE gene (−(ΔCT gene − ΔCT GAPDH )), normalized to mock infected cells are shown (n = 4; Mean ± SD). *P = 0.015 and ***P = 0.0003 (Holm-Sidak t test with α = 0.05).

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Mutagenesis, Sequencing, Infection, Expressing

    Persistently infected bat cells are resistant to super-infection with wildtype or ΔORF5 MERS-CoV. ( a–c ) Efk cells persistently infected with MERS-CoV were superinfected with W + MERS-CoV (blue) and transcript levels for ( a ) upE, ( b ) ORF5 or ( c ) E. fuscus dipeptidyl peptidase 4 (DPP4) were measured. The expression levels of upE, ORF5 and DPP4 transcripts (−(ΔCT gene − ΔCT GAPDH )) were also measured in Efk cells that were persistently infected with MERS-CoV in the absence of additional virus (red) and naïve Efk cells infected with W + MERS-CoV (green), with respect to time 0 for input W + virus (n = 3; Mean ± SD). DPP4 qRT-PCR amplicons were analyzed on an agarose gel (gel inset) and a ratio of basal DPP4 transcript levels (−(ΔCT DPP4 − ΔCT GAPDH )) in naïve, uninfected Efk and MRC5 cells is shown (right panel). ***P

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: Persistently infected bat cells are resistant to super-infection with wildtype or ΔORF5 MERS-CoV. ( a–c ) Efk cells persistently infected with MERS-CoV were superinfected with W + MERS-CoV (blue) and transcript levels for ( a ) upE, ( b ) ORF5 or ( c ) E. fuscus dipeptidyl peptidase 4 (DPP4) were measured. The expression levels of upE, ORF5 and DPP4 transcripts (−(ΔCT gene − ΔCT GAPDH )) were also measured in Efk cells that were persistently infected with MERS-CoV in the absence of additional virus (red) and naïve Efk cells infected with W + MERS-CoV (green), with respect to time 0 for input W + virus (n = 3; Mean ± SD). DPP4 qRT-PCR amplicons were analyzed on an agarose gel (gel inset) and a ratio of basal DPP4 transcript levels (−(ΔCT DPP4 − ΔCT GAPDH )) in naïve, uninfected Efk and MRC5 cells is shown (right panel). ***P

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Infection, Expressing, Quantitative RT-PCR, Agarose Gel Electrophoresis

    Bat cells can be persistently infected with MERS-CoV. ( a ) Big brown bat kidney cells (Efk) were infected with MERS-CoV (MOI = 0.01 TCID 50 /cell) for 12 days and then passaged weekly. Supernatant was collected during each passage to determine the presence of virus by titration on Vero cells, along with immunofluorescent and electron microscopic studies of infected cells. ( b ) Levels of MERS-CoV at different times following initial infection. ( c ) Phase contrast micrographs showing cytopathic effects on MERS-CoV infection and subsequent recovery of Efk cells at various time points. ( d ) Immunofluorescent images showing MERS-CoV nucleocapsid (N) protein in persistently infected Efk cells (bottom row; red arrows). The contrast for persistently infected Efk cells (inset) was adjusted to visualize low levels of protein. High MOI acute infection (middle row) and mock infection of Efk cells (top row) were used as positive and negative controls, respectively. Images were processed using ImageJ. ( e ) In-situ hybridization to detect the presence of MERS-CoV nucleoprotein RNA in persistently infected Efk cells. High, intermediate and low levels of MERS-CoV nucleoprotein RNA have been shown in the insets. Acutely infected (right) and mock infected (left) cells were used as positive and negative controls, respectively.

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: Bat cells can be persistently infected with MERS-CoV. ( a ) Big brown bat kidney cells (Efk) were infected with MERS-CoV (MOI = 0.01 TCID 50 /cell) for 12 days and then passaged weekly. Supernatant was collected during each passage to determine the presence of virus by titration on Vero cells, along with immunofluorescent and electron microscopic studies of infected cells. ( b ) Levels of MERS-CoV at different times following initial infection. ( c ) Phase contrast micrographs showing cytopathic effects on MERS-CoV infection and subsequent recovery of Efk cells at various time points. ( d ) Immunofluorescent images showing MERS-CoV nucleocapsid (N) protein in persistently infected Efk cells (bottom row; red arrows). The contrast for persistently infected Efk cells (inset) was adjusted to visualize low levels of protein. High MOI acute infection (middle row) and mock infection of Efk cells (top row) were used as positive and negative controls, respectively. Images were processed using ImageJ. ( e ) In-situ hybridization to detect the presence of MERS-CoV nucleoprotein RNA in persistently infected Efk cells. High, intermediate and low levels of MERS-CoV nucleoprotein RNA have been shown in the insets. Acutely infected (right) and mock infected (left) cells were used as positive and negative controls, respectively.

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Infection, Titration, In Situ Hybridization

    IRF3 and MAP kinase-mediated signaling regulate persistent infection in Efk cells. Persistently infected Efk cells were transfected with siRNA targeting IRF3 mRNA and the subsequent effect on virus replication was measured. ( a) MERS-CoV titres in persistently infected bat cells 24 hours post treatment with IRF3-siRNA (red bar; n = 4; Mean ± SD). Scrambled siRNA (blue bar; control-siRNA) was used as a negative control. **P = 0.0049 (Unpaired t test with α = 0.05). ( b) Western blot for IRF3 and GAPDH in Efk cells treated or mock treated with IRF3 siRNA. (c) MERS-CoV upE transcript levels in Efk cells after treatment or mock treatment with MAPK inhibitor, URMC-99 for 24 and 48 hours. * P = 0.0053 ( Holm-Sidak t test with α=0.05). ( d) RERG transcript levels 48-hours post treatment with URMC-99 (n = 4; Mean ± SD). * P = 0.017 ( Holm-Sidak t test with α = 0.05). For full size gel images in (b) , see supplementary Fig. S3 .

    Journal: Scientific Reports

    Article Title: Selection of viral variants during persistent infection of insectivorous bat cells with Middle East respiratory syndrome coronavirus

    doi: 10.1038/s41598-020-64264-1

    Figure Lengend Snippet: IRF3 and MAP kinase-mediated signaling regulate persistent infection in Efk cells. Persistently infected Efk cells were transfected with siRNA targeting IRF3 mRNA and the subsequent effect on virus replication was measured. ( a) MERS-CoV titres in persistently infected bat cells 24 hours post treatment with IRF3-siRNA (red bar; n = 4; Mean ± SD). Scrambled siRNA (blue bar; control-siRNA) was used as a negative control. **P = 0.0049 (Unpaired t test with α = 0.05). ( b) Western blot for IRF3 and GAPDH in Efk cells treated or mock treated with IRF3 siRNA. (c) MERS-CoV upE transcript levels in Efk cells after treatment or mock treatment with MAPK inhibitor, URMC-99 for 24 and 48 hours. * P = 0.0053 ( Holm-Sidak t test with α=0.05). ( d) RERG transcript levels 48-hours post treatment with URMC-99 (n = 4; Mean ± SD). * P = 0.017 ( Holm-Sidak t test with α = 0.05). For full size gel images in (b) , see supplementary Fig. S3 .

    Article Snippet: Primary staining for MERS-CoV nucleoprotein (N) and GAPDH was performed using 1:100 dilution of rabbit anti-MERS-CoV N (Sino Biological, Cat #40068-RP01) and mouse anti-GAPDH (EMD Milipore, Cat #AB2302).

    Techniques: Infection, Transfection, Negative Control, Western Blot

    IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.

    Journal: Vaccine

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    doi: 10.1016/j.vaccine.2016.11.064

    Figure Lengend Snippet: IHC detection of virus antigen expression in mouse tissue after challenge with MERS-CoV. Lung (A–C) and trachea (D–F) sections were assessed using rabbit polyclonal antibody to MERS-CoV nucleoprotein (NP) 3 days after the MERS-CoV challenge. The dark purple spot marked the inflammatory cell infiltration, and the brown particle marked the antigen of MERS-CoV. The MERS-CoV was located mainly in the trachea. Additionally, the lung tissue showed MERS-CoV expression in all immunized groups.

    Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

    Techniques: Immunohistochemistry, Expressing

    rNTD or rRBD vaccination reduced respiratory tract pathology in mice after MERS-CoV challenge. Representative results of hematoxylin-eosin (HE) staining in the lung (A – C) and trachea (D – F) of mock-treated or immunized mice. Severe lesions including the loss of pulmonary alveolus (represented by the white vacuole) and diffuse inflammatory cell infiltration (represented by the dark purple point) are shown (figure A). In contrast, milder lesions were observed among mice immunized with rRBD (figure B) or NTD (figure C), as the pulmonary alveolus was highly visible with less inflammatory cell infiltration. Inflammatory cell infiltration and impaired epithelium of the tunica mucosa bronchiorum were seen in the mock group (D). rRBD (E) or rNTD (F) alleviated the pathologic damage in the trachea of immunized mice. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: Vaccine

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    doi: 10.1016/j.vaccine.2016.11.064

    Figure Lengend Snippet: rNTD or rRBD vaccination reduced respiratory tract pathology in mice after MERS-CoV challenge. Representative results of hematoxylin-eosin (HE) staining in the lung (A – C) and trachea (D – F) of mock-treated or immunized mice. Severe lesions including the loss of pulmonary alveolus (represented by the white vacuole) and diffuse inflammatory cell infiltration (represented by the dark purple point) are shown (figure A). In contrast, milder lesions were observed among mice immunized with rRBD (figure B) or NTD (figure C), as the pulmonary alveolus was highly visible with less inflammatory cell infiltration. Inflammatory cell infiltration and impaired epithelium of the tunica mucosa bronchiorum were seen in the mock group (D). rRBD (E) or rNTD (F) alleviated the pathologic damage in the trachea of immunized mice. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

    Techniques: Mouse Assay, Staining

    Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Journal: Vaccine

    Article Title: The recombinant N-terminal domain of spike proteins is a potential vaccine against Middle East respiratory syndrome coronavirus (MERS-CoV) infection

    doi: 10.1016/j.vaccine.2016.11.064

    Figure Lengend Snippet: Description of the N-terminal domain (NTD) immunogen and vaccination schedule. (A) The location of the NTD protein on the Middle East respiratory syndrome coronavirus MERS-CoV spike (S) protein. The recombinant (r)NTD protein consists of 336 amino acid (aa) residues (18–353) of S protein. A gp67 signal peptide (SP) was added to the N terminus for expression of the rNTD protein. (B) Purified rNTD protein detected by SDS-PAGE (left) and Western blot (right). The purified rNTD protein was separated by a 10% SDS-PAGE and stained with 0.25% Coomassie brilliant blue. Anti-NTD polyclonal antibody and infrared ray-labeled secondary antibody were used for the Western blot assay. Lane 1: protein molecular weight marker; lane 2: purified rNTD protein. (C). Vaccination schedule and detection. Mice received three vaccinations consisting of 5 or 10 μg of rNTD protein combined with adjuvants at 4-week intervals. Sera were collected at the indicated times to analyze the humoral immune response. Six mice from each group were sacrificed 2 weeks after the last immunization. The spleens were harvested for enzyme-linked immunospot (ELISpot), intracellular cytokine staining (ICS), and cytometric bead array (CBA) assays. In parallel experiments, the remaining mice were challenged with MERS-CoV to detect the protective effect elicited by the rNTD protein. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: Rabbit-serum-derived polyclonal antibody against nucleoprotein (cat: 100213-RP02; Sino Biological Inc., Beijing, CHN) was incubated with the sections at 1:1000 dilution; goat anti-rabbit (cat: pv-9001; ZSGB-Bio, Beijing, CHN) secondary antibody was used at 1:2000, and sections were evaluated using light microscopy.

    Techniques: Recombinant, Expressing, Purification, SDS Page, Western Blot, Staining, Labeling, Molecular Weight, Marker, Mouse Assay, Enzyme-linked Immunospot, Crocin Bleaching Assay