biotinylated pig ifn α R&D Systems Search Results


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  • 95
    Millipore ifn γ
    Ifn γ, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 4538 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories avidin biotin blocking kit
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    R&D Systems biotinylated pig ifn α
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
    Biotinylated Pig Ifn α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PBL Assay ifn α
    <t>IFN-α</t> can enhance autoreactive B cell responses to non-optimal DNA fragment ICs
    Ifn α, supplied by PBL Assay, used in various techniques. Bioz Stars score: 99/100, based on 1090 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PBL Biomedical Laboratories ifn α
    Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated <t>anti-IFN-α</t> antibody.
    Ifn α, supplied by PBL Biomedical Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 769 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems anti porcine ifn α
    Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated <t>anti-IFN-α</t> antibody.
    Anti Porcine Ifn α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 84/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems biotinylated anti porcine ifn α
    Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated <t>anti-IFN-α</t> antibody.
    Biotinylated Anti Porcine Ifn α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems ifn α protein
    Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated <t>anti-IFN-α</t> antibody.
    Ifn α Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Mab Technologies ifn alpha
    Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated <t>anti-IFN-α</t> antibody.
    Ifn Alpha, supplied by Mab Technologies, used in various techniques. Bioz Stars score: 99/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems mouse mab against porcine ifn α
    Investigation of CSFV-infected pDC-derived cytokines on the MHC class II expression of NK and γδ T cells. (A) pDCs or mDCs were exposed to CSFV C-strain, Brescia strain, or a mock virus-infected cryolysate supernatant, and after 24 h in culture, the levels of TNF-α and type I <t>IFN</t> in the supernatants were analyzed using a porcine cytokine multiplex ELISA and Mx/CAT reporter gene assay, respectively. (B) Enriched NK and γδ T cells were then cultured in cRPMI medium alone or in the presence of porcine recombinant <t>IFN-α</t> and/or TNF-α, at concentrations comparable to those detected in infected pDC cocultures, and the expression of MHC-II was assessed by flow cytometry after 24 h. The mean ± SEM data from three independent experiments utilizing different animals are shown. The values for each group were compared to the corresponding unstimulated control (mock stimulation in panel A and no cytokine treatment in panel B) using a one-way ANOVA, followed by Dunnett's multiple comparison test: ***, P
    Mouse Mab Against Porcine Ifn α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 77/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems mouse ifn alpha beta r2 biotinylated antibody
    Investigation of CSFV-infected pDC-derived cytokines on the MHC class II expression of NK and γδ T cells. (A) pDCs or mDCs were exposed to CSFV C-strain, Brescia strain, or a mock virus-infected cryolysate supernatant, and after 24 h in culture, the levels of TNF-α and type I <t>IFN</t> in the supernatants were analyzed using a porcine cytokine multiplex ELISA and Mx/CAT reporter gene assay, respectively. (B) Enriched NK and γδ T cells were then cultured in cRPMI medium alone or in the presence of porcine recombinant <t>IFN-α</t> and/or TNF-α, at concentrations comparable to those detected in infected pDC cocultures, and the expression of MHC-II was assessed by flow cytometry after 24 h. The mean ± SEM data from three independent experiments utilizing different animals are shown. The values for each group were compared to the corresponding unstimulated control (mock stimulation in panel A and no cytokine treatment in panel B) using a one-way ANOVA, followed by Dunnett's multiple comparison test: ***, P
    Mouse Ifn Alpha Beta R2 Biotinylated Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Miltenyi Biotec ifn α
    Investigation of CSFV-infected pDC-derived cytokines on the MHC class II expression of NK and γδ T cells. (A) pDCs or mDCs were exposed to CSFV C-strain, Brescia strain, or a mock virus-infected cryolysate supernatant, and after 24 h in culture, the levels of TNF-α and type I <t>IFN</t> in the supernatants were analyzed using a porcine cytokine multiplex ELISA and Mx/CAT reporter gene assay, respectively. (B) Enriched NK and γδ T cells were then cultured in cRPMI medium alone or in the presence of porcine recombinant <t>IFN-α</t> and/or TNF-α, at concentrations comparable to those detected in infected pDC cocultures, and the expression of MHC-II was assessed by flow cytometry after 24 h. The mean ± SEM data from three independent experiments utilizing different animals are shown. The values for each group were compared to the corresponding unstimulated control (mock stimulation in panel A and no cytokine treatment in panel B) using a one-way ANOVA, followed by Dunnett's multiple comparison test: ***, P
    Ifn α, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 99/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson biotinylated α ifn γ ab
    Plasma concentrations of cytokines. Plasma concentrations of <t>IFN-γ</t> ( A ), IL-6 ( B ), IL-1β ( C ), IL-5 ( D ), TNF-α ( E ), and IL-2 ( F ) were measured for six animals in the SEB negative control, PBS positive control, and SEB-treated groups
    Biotinylated α Ifn γ Ab, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 80/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems ifn α
    Diarrhoea scores, percentage of piglets with jejunal pathology, serum interferon-α <t>(IFN-α)</t> and infectious virus shed after 10 focus-forming units (ffu) or 1000 ffu of human rotavirus (HRV). Diarrhoea scores (a), percentage of piglets with jejunal pathology (b), serum IFN-α (pg/ml, c) and infectious virus shedding (ffu/ml, d) are shown after 10 ffu (○) and 1000 ffu (▪) from post-infection day (PID) 0–6 ( x -axis). The number of piglets studied and the percentage of piglets that developed diarrhoea, shedding and jejunal pathology at each time-point is shown in each graph. Data-points with an asterisk differ significantly between doses at each time-point. The error bars denote the standard error of the mean (SEM) and values with an asterisk denote significant differences between groups at each PID. The HRV faecal shedding and diarrhoea scores were analysed using analysis of variance followed by Duncan's rank sum test, the percentage of pigs with diarrhoea and RV shedding was analysed using Fisher's exact test. Serum IFN-α was analysed using Kruskall–Wallis rank sum test. A P
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    Becton Dickinson α ifn γ biotinylated mab
    Diarrhoea scores, percentage of piglets with jejunal pathology, serum interferon-α <t>(IFN-α)</t> and infectious virus shed after 10 focus-forming units (ffu) or 1000 ffu of human rotavirus (HRV). Diarrhoea scores (a), percentage of piglets with jejunal pathology (b), serum IFN-α (pg/ml, c) and infectious virus shedding (ffu/ml, d) are shown after 10 ffu (○) and 1000 ffu (▪) from post-infection day (PID) 0–6 ( x -axis). The number of piglets studied and the percentage of piglets that developed diarrhoea, shedding and jejunal pathology at each time-point is shown in each graph. Data-points with an asterisk differ significantly between doses at each time-point. The error bars denote the standard error of the mean (SEM) and values with an asterisk denote significant differences between groups at each PID. The HRV faecal shedding and diarrhoea scores were analysed using analysis of variance followed by Duncan's rank sum test, the percentage of pigs with diarrhoea and RV shedding was analysed using Fisher's exact test. Serum IFN-α was analysed using Kruskall–Wallis rank sum test. A P
    α Ifn γ Biotinylated Mab, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems biotinylated goat anti mouse ifn α βr1 antibody
    Diarrhoea scores, percentage of piglets with jejunal pathology, serum interferon-α <t>(IFN-α)</t> and infectious virus shed after 10 focus-forming units (ffu) or 1000 ffu of human rotavirus (HRV). Diarrhoea scores (a), percentage of piglets with jejunal pathology (b), serum IFN-α (pg/ml, c) and infectious virus shedding (ffu/ml, d) are shown after 10 ffu (○) and 1000 ffu (▪) from post-infection day (PID) 0–6 ( x -axis). The number of piglets studied and the percentage of piglets that developed diarrhoea, shedding and jejunal pathology at each time-point is shown in each graph. Data-points with an asterisk differ significantly between doses at each time-point. The error bars denote the standard error of the mean (SEM) and values with an asterisk denote significant differences between groups at each PID. The HRV faecal shedding and diarrhoea scores were analysed using analysis of variance followed by Duncan's rank sum test, the percentage of pigs with diarrhoea and RV shedding was analysed using Fisher's exact test. Serum IFN-α was analysed using Kruskall–Wallis rank sum test. A P
    Biotinylated Goat Anti Mouse Ifn α βr1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems biotinylated goat anti human ifn γ
    Analysis of cytokine release from healthy human T-cells. ( A , B ) T-cells captured on 300 μm diameter anti-CD4 Ab spots next to cytokine-sensing Ab microarray. Cells were mitogenically activated on-chip for 4h. Cytokine signal was developed by incubating microarrays with HRP-labeled anti-cytokine Abs and color reagent (DAB). ( C ) Relative cytokine signal intensity determined by lensfree on-chip imaging of <t>IFN-γ,</t> TNF-α and IL-2-sensing spots. Cytokine signal was observed only when T-cells were mitogenically stimulated with PMA/Ionomyocin. In a parallel experiment, Ab microarrays were exposed to a known reference concentration (100 ng/ml) of recombinant IFN-γ, TNF-α and IL-2. This reference signal was used to estimate the amount of secreted cytokines.
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    Quanterix ifn α
    Analysis of cytokine release from healthy human T-cells. ( A , B ) T-cells captured on 300 μm diameter anti-CD4 Ab spots next to cytokine-sensing Ab microarray. Cells were mitogenically activated on-chip for 4h. Cytokine signal was developed by incubating microarrays with HRP-labeled anti-cytokine Abs and color reagent (DAB). ( C ) Relative cytokine signal intensity determined by lensfree on-chip imaging of <t>IFN-γ,</t> TNF-α and IL-2-sensing spots. Cytokine signal was observed only when T-cells were mitogenically stimulated with PMA/Ionomyocin. In a parallel experiment, Ab microarrays were exposed to a known reference concentration (100 ng/ml) of recombinant IFN-γ, TNF-α and IL-2. This reference signal was used to estimate the amount of secreted cytokines.
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    PBL Assay recombinant porcine ifn α
    Serum cytokine levels in PRRSV-infected pigs. Sera collected from pigs (n = 5) at 0, 3, 7, 14, 21 and 28 dpc were analysed for a <t>IFN-α;</t> b TNF-α; c IL-12; d IL-10; and e IL-4 by ELISA. Each bar represents the average cytokine amount from 5 pigs ± SEM. Bars showing different letters represent values that differ significantly from each other ( p
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    R&D Systems anti pig ifn α
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
    Anti Pig Ifn α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 83/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human il 29 ifn lambda 1 biotinylated antibody
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    R&D Systems recombinant mouse ifn α
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    Thermo Fisher biotinylated α ifn γ mab
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    Agilent technologies avidin biotin complex
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    Mab Technologies biotinylated α mouse ifn γ
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    Agilent technologies avidin biotin peroxidase complex abc
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    PBL Biomedical Laboratories ifn α recombinant protein
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
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    R&D Systems anti mouse ifn α antibody
    Evolution of <t>IFN-α</t> levels (Limit of detection: 3.9 pg/mL)
    Anti Mouse Ifn α Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Evolution of IFN-α levels (Limit of detection: 3.9 pg/mL)

    Journal: Porcine Health Management

    Article Title: Immune response development after vaccination of 1-day-old naïve pigs with a Porcine Reproductive and Respiratory Syndrome 1-based modified live virus vaccine

    doi: 10.1186/s40813-018-0112-7

    Figure Lengend Snippet: Evolution of IFN-α levels (Limit of detection: 3.9 pg/mL)

    Article Snippet: Briefly, plates were coated with anti-pig IFN-α (mAb K9, R & D systems) at 1.1 Ug/mL; 50 μl of each sample and biotinylated pig IFN-α (mAb F17, R & D systems) were added.

    Techniques:

    IFN-α can enhance autoreactive B cell responses to non-optimal DNA fragment ICs

    Journal:

    Article Title: Autoreactive B Cells Discriminate CpG-rich and CpG-poor DNA and This Response is Modulated by IFN-? 1

    doi:

    Figure Lengend Snippet: IFN-α can enhance autoreactive B cell responses to non-optimal DNA fragment ICs

    Article Snippet: In some experiments, cells were primed with IFN-α (PBL InterferonSource, 1000 U/ml) prior to addition of stimuli.

    Techniques:

    Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated anti-IFN-α antibody.

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Characterization of Virus-Responsive Plasmacytoid Dendritic Cells in the Rhesus Macaque

    doi: 10.1128/CDLI.12.3.426-435.2005

    Figure Lengend Snippet: Microscopic analysis of macaque PDC. (A) Macaque PDC were enriched from PBMC by negative selection and then Giemsa stained and analyzed for morphology, revealing cells with characteristic PDC morphology. Positively selected PDC were mock (B) or HSV (C) stimulated for 6 h and then stained with Alexa Fluor 680-conjugated anti-IFN-α antibody.

    Article Snippet: The following day, cells were washed twice with PBS-2% FCS, permeabilized with 0.5% saponin (Sigma-Aldrich) in PBS-2% FCS for 30 min at room temperature, and then incubated with 50 ng of biotinylated 293 monoclonal antibody (MAb) to IFN-α (obtained from G. V. Alm, Uppsala, Sweden) or a commercially available antibody to IFN-α (clone MMHA-2, PBL Biomedical Laboratories, Piscataway, N.J.).

    Techniques: Selection, Staining

    Intracellular flow cytometric detection of IFN-α production by HSV-stimulated macaque and human PDC. (A) Human PBMC (left panel) were gated as CD123 + HLA-DR + cells and then further gated for IFN-α production to determine the percentage of PDC that produce IFN-α in response to mock and HSV stimulation. The numbers in the upper right quadrants are the percentages of IFN-α + PDC. (B) Macaque PBMC were gated as described for human PDC in panel A to determine the percentage of PDC producing IFN-α in response to mock and HSV stimulation. The data shown are representative experiments of 11 human samples and 28 macaque samples. (C) The percentage of PDC producing IFN-α ranged from 12 to 80% in humans and 36 to 82% in macaques. (D) There was no significant difference in the percentages of PDC producing IFN-α between freshly drawn macaque blood and overnight-shipped blood, where PBMC derived from fresh and shipped blood are shown with the same symbol for an individual animal. The boxes in panels C and D represent the upper and lower quartiles, with the medians shown inside the boxes. The lines extend out to either the upper quartiles plus 1.5 times the interquartile range or the lower quartiles minus 1.5 times the interquartile range.

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Characterization of Virus-Responsive Plasmacytoid Dendritic Cells in the Rhesus Macaque

    doi: 10.1128/CDLI.12.3.426-435.2005

    Figure Lengend Snippet: Intracellular flow cytometric detection of IFN-α production by HSV-stimulated macaque and human PDC. (A) Human PBMC (left panel) were gated as CD123 + HLA-DR + cells and then further gated for IFN-α production to determine the percentage of PDC that produce IFN-α in response to mock and HSV stimulation. The numbers in the upper right quadrants are the percentages of IFN-α + PDC. (B) Macaque PBMC were gated as described for human PDC in panel A to determine the percentage of PDC producing IFN-α in response to mock and HSV stimulation. The data shown are representative experiments of 11 human samples and 28 macaque samples. (C) The percentage of PDC producing IFN-α ranged from 12 to 80% in humans and 36 to 82% in macaques. (D) There was no significant difference in the percentages of PDC producing IFN-α between freshly drawn macaque blood and overnight-shipped blood, where PBMC derived from fresh and shipped blood are shown with the same symbol for an individual animal. The boxes in panels C and D represent the upper and lower quartiles, with the medians shown inside the boxes. The lines extend out to either the upper quartiles plus 1.5 times the interquartile range or the lower quartiles minus 1.5 times the interquartile range.

    Article Snippet: The following day, cells were washed twice with PBS-2% FCS, permeabilized with 0.5% saponin (Sigma-Aldrich) in PBS-2% FCS for 30 min at room temperature, and then incubated with 50 ng of biotinylated 293 monoclonal antibody (MAb) to IFN-α (obtained from G. V. Alm, Uppsala, Sweden) or a commercially available antibody to IFN-α (clone MMHA-2, PBL Biomedical Laboratories, Piscataway, N.J.).

    Techniques: Flow Cytometry, Derivative Assay

    Frequency of IFN-α-producing cells in macaque and human PBMC. Macaque and human PBMC were either mock stimulated (A and B), HSV stimulated (A), or SV stimulated (B) for 6 h. The frequency of IFN-α-producing cells was determined by ELISPOT assay. The mean values for 6 human samples and 12 macaque samples are shown. The error bars represent one standard error of the mean. The number of IPC detected in macaque PBMC in response to HSV and SV was significantly lower than in humans ( P

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Characterization of Virus-Responsive Plasmacytoid Dendritic Cells in the Rhesus Macaque

    doi: 10.1128/CDLI.12.3.426-435.2005

    Figure Lengend Snippet: Frequency of IFN-α-producing cells in macaque and human PBMC. Macaque and human PBMC were either mock stimulated (A and B), HSV stimulated (A), or SV stimulated (B) for 6 h. The frequency of IFN-α-producing cells was determined by ELISPOT assay. The mean values for 6 human samples and 12 macaque samples are shown. The error bars represent one standard error of the mean. The number of IPC detected in macaque PBMC in response to HSV and SV was significantly lower than in humans ( P

    Article Snippet: The following day, cells were washed twice with PBS-2% FCS, permeabilized with 0.5% saponin (Sigma-Aldrich) in PBS-2% FCS for 30 min at room temperature, and then incubated with 50 ng of biotinylated 293 monoclonal antibody (MAb) to IFN-α (obtained from G. V. Alm, Uppsala, Sweden) or a commercially available antibody to IFN-α (clone MMHA-2, PBL Biomedical Laboratories, Piscataway, N.J.).

    Techniques: Enzyme-linked Immunospot

    PDC are the predominant IPC in HSV-stimulated macaque PBMC. PBMC were either mock (A) or HSV (B) stimulated and gated based on forward scatter and intracellular expression of IFN-α. (C) Of the cells that are IFN-α + , 90% are HLA-DR + and CD123 + . The data are shown for one representative experiment of two.

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Characterization of Virus-Responsive Plasmacytoid Dendritic Cells in the Rhesus Macaque

    doi: 10.1128/CDLI.12.3.426-435.2005

    Figure Lengend Snippet: PDC are the predominant IPC in HSV-stimulated macaque PBMC. PBMC were either mock (A) or HSV (B) stimulated and gated based on forward scatter and intracellular expression of IFN-α. (C) Of the cells that are IFN-α + , 90% are HLA-DR + and CD123 + . The data are shown for one representative experiment of two.

    Article Snippet: The following day, cells were washed twice with PBS-2% FCS, permeabilized with 0.5% saponin (Sigma-Aldrich) in PBS-2% FCS for 30 min at room temperature, and then incubated with 50 ng of biotinylated 293 monoclonal antibody (MAb) to IFN-α (obtained from G. V. Alm, Uppsala, Sweden) or a commercially available antibody to IFN-α (clone MMHA-2, PBL Biomedical Laboratories, Piscataway, N.J.).

    Techniques: Expressing

    Investigation of CSFV-infected pDC-derived cytokines on the MHC class II expression of NK and γδ T cells. (A) pDCs or mDCs were exposed to CSFV C-strain, Brescia strain, or a mock virus-infected cryolysate supernatant, and after 24 h in culture, the levels of TNF-α and type I IFN in the supernatants were analyzed using a porcine cytokine multiplex ELISA and Mx/CAT reporter gene assay, respectively. (B) Enriched NK and γδ T cells were then cultured in cRPMI medium alone or in the presence of porcine recombinant IFN-α and/or TNF-α, at concentrations comparable to those detected in infected pDC cocultures, and the expression of MHC-II was assessed by flow cytometry after 24 h. The mean ± SEM data from three independent experiments utilizing different animals are shown. The values for each group were compared to the corresponding unstimulated control (mock stimulation in panel A and no cytokine treatment in panel B) using a one-way ANOVA, followed by Dunnett's multiple comparison test: ***, P

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Partial Activation of Natural Killer and γδ T Cells by Classical Swine Fever Viruses Is Associated with Type I Interferon Elicited from Plasmacytoid Dendritic Cells

    doi: 10.1128/CVI.00382-14

    Figure Lengend Snippet: Investigation of CSFV-infected pDC-derived cytokines on the MHC class II expression of NK and γδ T cells. (A) pDCs or mDCs were exposed to CSFV C-strain, Brescia strain, or a mock virus-infected cryolysate supernatant, and after 24 h in culture, the levels of TNF-α and type I IFN in the supernatants were analyzed using a porcine cytokine multiplex ELISA and Mx/CAT reporter gene assay, respectively. (B) Enriched NK and γδ T cells were then cultured in cRPMI medium alone or in the presence of porcine recombinant IFN-α and/or TNF-α, at concentrations comparable to those detected in infected pDC cocultures, and the expression of MHC-II was assessed by flow cytometry after 24 h. The mean ± SEM data from three independent experiments utilizing different animals are shown. The values for each group were compared to the corresponding unstimulated control (mock stimulation in panel A and no cytokine treatment in panel B) using a one-way ANOVA, followed by Dunnett's multiple comparison test: ***, P

    Article Snippet: A mouse MAb against porcine IFN-α (clone F17; R & D Systems) was added at 1.82 μg/ml and sections incubated at 4°C for 18 to 20 h. The sections were washed in TBST and incubated for 30 min with a 1:100 dilution of biotinylated secondary antibody (biotin-SP-conjugated AffiniPure Fab fragment goat anti-mouse IgG [H+L]; Jackson ImmunoResearch, Stratech, Newmarket, United Kingdom), and normal goat serum (1:66 dilution) and normal pig serum (1:33 dilution) were added before being washed twice in TBST.

    Techniques: Infection, Derivative Assay, Expressing, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Reporter Gene Assay, Cell Culture, Recombinant, Flow Cytometry, Cytometry

    Quantification of local and systemic type I IFN following C-strain vaccination and/or virulent CSFV Brescia challenge. Serum and tonsils were collected longitudinally following C-strain vaccination and challenge 5 days later with CSFV Brescia strain. Cells expressing IFN-α were enumerated in the interfollicular areas (closed symbols) and follicles (open symbols) of the tonsils by immunohistochemistry (A), and the levels of type I IFN in serum were determined using an Mx/CAT reporter gene assay (B). The data represent the mean of 3 pigs/group ± SEM. Statistical analyses were performed using a one-way ANOVA, followed by Bonferroni's multiple comparison test: ***, P

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Partial Activation of Natural Killer and γδ T Cells by Classical Swine Fever Viruses Is Associated with Type I Interferon Elicited from Plasmacytoid Dendritic Cells

    doi: 10.1128/CVI.00382-14

    Figure Lengend Snippet: Quantification of local and systemic type I IFN following C-strain vaccination and/or virulent CSFV Brescia challenge. Serum and tonsils were collected longitudinally following C-strain vaccination and challenge 5 days later with CSFV Brescia strain. Cells expressing IFN-α were enumerated in the interfollicular areas (closed symbols) and follicles (open symbols) of the tonsils by immunohistochemistry (A), and the levels of type I IFN in serum were determined using an Mx/CAT reporter gene assay (B). The data represent the mean of 3 pigs/group ± SEM. Statistical analyses were performed using a one-way ANOVA, followed by Bonferroni's multiple comparison test: ***, P

    Article Snippet: A mouse MAb against porcine IFN-α (clone F17; R & D Systems) was added at 1.82 μg/ml and sections incubated at 4°C for 18 to 20 h. The sections were washed in TBST and incubated for 30 min with a 1:100 dilution of biotinylated secondary antibody (biotin-SP-conjugated AffiniPure Fab fragment goat anti-mouse IgG [H+L]; Jackson ImmunoResearch, Stratech, Newmarket, United Kingdom), and normal goat serum (1:66 dilution) and normal pig serum (1:33 dilution) were added before being washed twice in TBST.

    Techniques: Expressing, Immunohistochemistry, Reporter Gene Assay

    Plasma concentrations of cytokines. Plasma concentrations of IFN-γ ( A ), IL-6 ( B ), IL-1β ( C ), IL-5 ( D ), TNF-α ( E ), and IL-2 ( F ) were measured for six animals in the SEB negative control, PBS positive control, and SEB-treated groups

    Journal: The Journal of Immunology Author Choice

    Article Title: Interference of the T Cell and Antigen-Presenting Cell Costimulatory Pathway Using CTLA4-Ig (Abatacept) Prevents Staphylococcal Enterotoxin B Pathology

    doi: 10.4049/jimmunol.1601525

    Figure Lengend Snippet: Plasma concentrations of cytokines. Plasma concentrations of IFN-γ ( A ), IL-6 ( B ), IL-1β ( C ), IL-5 ( D ), TNF-α ( E ), and IL-2 ( F ) were measured for six animals in the SEB negative control, PBS positive control, and SEB-treated groups

    Article Snippet: Plates were blocked with 1% BSA; then 50 μl of the culture supernatant or the relevant WHO International Standards (Second WHO Internal Standard for TNF-α NIBSC 88/786, IL-2 standard NIBSC 86/564, IFN-γ standard NIBSC 88/606) was added to the plates together with 50 μl of biotinylated anti–TNF-α (clone H91; NIBSC), or biotinylated anti–IL-2 (R & D BAF202) or biotinylated anti–IFN-γ (BD 554550; BD Biosciences).

    Techniques: Negative Control, Positive Control

    Lung tissue extract concentrations of IFN-γ, IL-6, CCL2, IL-1β, IL-5, and TNF-α. Lung tissue extract concentrations of IFN-γ ( A ), IL-6 ( B ), CCL2 ( C ), IL-1β ( D ), IL-5 ( E ), and TNF-α ( F ) were measured. One-way

    Journal: The Journal of Immunology Author Choice

    Article Title: Interference of the T Cell and Antigen-Presenting Cell Costimulatory Pathway Using CTLA4-Ig (Abatacept) Prevents Staphylococcal Enterotoxin B Pathology

    doi: 10.4049/jimmunol.1601525

    Figure Lengend Snippet: Lung tissue extract concentrations of IFN-γ, IL-6, CCL2, IL-1β, IL-5, and TNF-α. Lung tissue extract concentrations of IFN-γ ( A ), IL-6 ( B ), CCL2 ( C ), IL-1β ( D ), IL-5 ( E ), and TNF-α ( F ) were measured. One-way

    Article Snippet: Plates were blocked with 1% BSA; then 50 μl of the culture supernatant or the relevant WHO International Standards (Second WHO Internal Standard for TNF-α NIBSC 88/786, IL-2 standard NIBSC 86/564, IFN-γ standard NIBSC 88/606) was added to the plates together with 50 μl of biotinylated anti–TNF-α (clone H91; NIBSC), or biotinylated anti–IL-2 (R & D BAF202) or biotinylated anti–IFN-γ (BD 554550; BD Biosciences).

    Techniques:

    Diarrhoea scores, percentage of piglets with jejunal pathology, serum interferon-α (IFN-α) and infectious virus shed after 10 focus-forming units (ffu) or 1000 ffu of human rotavirus (HRV). Diarrhoea scores (a), percentage of piglets with jejunal pathology (b), serum IFN-α (pg/ml, c) and infectious virus shedding (ffu/ml, d) are shown after 10 ffu (○) and 1000 ffu (▪) from post-infection day (PID) 0–6 ( x -axis). The number of piglets studied and the percentage of piglets that developed diarrhoea, shedding and jejunal pathology at each time-point is shown in each graph. Data-points with an asterisk differ significantly between doses at each time-point. The error bars denote the standard error of the mean (SEM) and values with an asterisk denote significant differences between groups at each PID. The HRV faecal shedding and diarrhoea scores were analysed using analysis of variance followed by Duncan's rank sum test, the percentage of pigs with diarrhoea and RV shedding was analysed using Fisher's exact test. Serum IFN-α was analysed using Kruskall–Wallis rank sum test. A P

    Journal: Immunology

    Article Title: Innate immune responses to human rotavirus in the neonatal gnotobiotic piglet disease model

    doi: 10.1111/j.1365-2567.2010.03298.x

    Figure Lengend Snippet: Diarrhoea scores, percentage of piglets with jejunal pathology, serum interferon-α (IFN-α) and infectious virus shed after 10 focus-forming units (ffu) or 1000 ffu of human rotavirus (HRV). Diarrhoea scores (a), percentage of piglets with jejunal pathology (b), serum IFN-α (pg/ml, c) and infectious virus shedding (ffu/ml, d) are shown after 10 ffu (○) and 1000 ffu (▪) from post-infection day (PID) 0–6 ( x -axis). The number of piglets studied and the percentage of piglets that developed diarrhoea, shedding and jejunal pathology at each time-point is shown in each graph. Data-points with an asterisk differ significantly between doses at each time-point. The error bars denote the standard error of the mean (SEM) and values with an asterisk denote significant differences between groups at each PID. The HRV faecal shedding and diarrhoea scores were analysed using analysis of variance followed by Duncan's rank sum test, the percentage of pigs with diarrhoea and RV shedding was analysed using Fisher's exact test. Serum IFN-α was analysed using Kruskall–Wallis rank sum test. A P

    Article Snippet: Biotinylated mAbs to IL-12 ,TNF-α, IL-6, IL-10 and IFN-α (R & D Systems) were diluted and added for a 2-hr incubation at room temperature.

    Techniques: Infection

    Correlation coefficient between interferon-α (IFN-α) positive plasmacytoid dendritic cells (pDCs) and serum IFN-α. A Spearman's correlation coefficient was calculated between IFN-α + pDCs and serum IFN-α. The R, P and number of piglets included in the analysis are shown in the graph.

    Journal: Immunology

    Article Title: Innate immune responses to human rotavirus in the neonatal gnotobiotic piglet disease model

    doi: 10.1111/j.1365-2567.2010.03298.x

    Figure Lengend Snippet: Correlation coefficient between interferon-α (IFN-α) positive plasmacytoid dendritic cells (pDCs) and serum IFN-α. A Spearman's correlation coefficient was calculated between IFN-α + pDCs and serum IFN-α. The R, P and number of piglets included in the analysis are shown in the graph.

    Article Snippet: Biotinylated mAbs to IL-12 ,TNF-α, IL-6, IL-10 and IFN-α (R & D Systems) were diluted and added for a 2-hr incubation at room temperature.

    Techniques:

    Intestinal frequencies of interferon-α (IFN-α) positive plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs) and macrophages/monocytes after human rotavirus (HRV) infection. (a) HRV inoculated and (b) medium alone inoculated representative dot plots of SWC3 versus CD4 at post-infection days (PID) 2, 4 and 7 are shown. Gates on SWC3 low CD4 + DCs (pDCs), SWC3 high CD4 − DCs (macrophage/monocytes) and SWC3 low CD4 − DCs (cDCs) and plots on each gate show IFN-α + DCs. Mean frequencies of IFN-α + pDCs (c), IFN-α + cDCs (d) and IFN-α + macrophages/monocytes (e) in piglets that received HRV (○) or medium alone (▪) are summarized. For each experiment 100 000 events were acquired by the flow cytometer. The number of piglets studied at each PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and bars with an asterisk differ significantly from the controls at each PID (Kruskall–Wallis rank sum test P ≤ 0.05).

    Journal: Immunology

    Article Title: Innate immune responses to human rotavirus in the neonatal gnotobiotic piglet disease model

    doi: 10.1111/j.1365-2567.2010.03298.x

    Figure Lengend Snippet: Intestinal frequencies of interferon-α (IFN-α) positive plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs) and macrophages/monocytes after human rotavirus (HRV) infection. (a) HRV inoculated and (b) medium alone inoculated representative dot plots of SWC3 versus CD4 at post-infection days (PID) 2, 4 and 7 are shown. Gates on SWC3 low CD4 + DCs (pDCs), SWC3 high CD4 − DCs (macrophage/monocytes) and SWC3 low CD4 − DCs (cDCs) and plots on each gate show IFN-α + DCs. Mean frequencies of IFN-α + pDCs (c), IFN-α + cDCs (d) and IFN-α + macrophages/monocytes (e) in piglets that received HRV (○) or medium alone (▪) are summarized. For each experiment 100 000 events were acquired by the flow cytometer. The number of piglets studied at each PID is shown at the bottom of each graph. The error bars denote the standard error of the mean (SEM) and bars with an asterisk differ significantly from the controls at each PID (Kruskall–Wallis rank sum test P ≤ 0.05).

    Article Snippet: Biotinylated mAbs to IL-12 ,TNF-α, IL-6, IL-10 and IFN-α (R & D Systems) were diluted and added for a 2-hr incubation at room temperature.

    Techniques: Infection, Flow Cytometry, Cytometry

    Analysis of cytokine release from healthy human T-cells. ( A , B ) T-cells captured on 300 μm diameter anti-CD4 Ab spots next to cytokine-sensing Ab microarray. Cells were mitogenically activated on-chip for 4h. Cytokine signal was developed by incubating microarrays with HRP-labeled anti-cytokine Abs and color reagent (DAB). ( C ) Relative cytokine signal intensity determined by lensfree on-chip imaging of IFN-γ, TNF-α and IL-2-sensing spots. Cytokine signal was observed only when T-cells were mitogenically stimulated with PMA/Ionomyocin. In a parallel experiment, Ab microarrays were exposed to a known reference concentration (100 ng/ml) of recombinant IFN-γ, TNF-α and IL-2. This reference signal was used to estimate the amount of secreted cytokines.

    Journal: Analytical chemistry

    Article Title: Lensfree Holographic Imaging of Antibody Microarrays for High-Throughput Detection of Leukocyte Numbers and Function

    doi: 10.1021/ac100142a

    Figure Lengend Snippet: Analysis of cytokine release from healthy human T-cells. ( A , B ) T-cells captured on 300 μm diameter anti-CD4 Ab spots next to cytokine-sensing Ab microarray. Cells were mitogenically activated on-chip for 4h. Cytokine signal was developed by incubating microarrays with HRP-labeled anti-cytokine Abs and color reagent (DAB). ( C ) Relative cytokine signal intensity determined by lensfree on-chip imaging of IFN-γ, TNF-α and IL-2-sensing spots. Cytokine signal was observed only when T-cells were mitogenically stimulated with PMA/Ionomyocin. In a parallel experiment, Ab microarrays were exposed to a known reference concentration (100 ng/ml) of recombinant IFN-γ, TNF-α and IL-2. This reference signal was used to estimate the amount of secreted cytokines.

    Article Snippet: Purified mouse anti-human IFN-γ Ab (clone K3.53), anti-IL2 Ab (clone 5355), anti-TNF-α/TNFSF1A Ab (clone 28401), biotinylated goat anti-human IFN-γ, IL2 and TNF-α/TNFSF1A Abs from R & D Systems (Minneapolis, MN).

    Techniques: Microarray, Chromatin Immunoprecipitation, Labeling, Imaging, Concentration Assay, Recombinant

    ( A ) Design of an antibody (Ab) microarray for capturing leukocytes and detecting secreted cytokines. T-cell Ab spots were print next to spots for detection of IFN-γ, TNF-α and IL-2. Anti-mouse IgG spots provided negative control whereas biotin spots were used for positive control. ( B ) Principle of operation of the assay. T-cells are captured in the immediate vicinity of cytokine-sensing Ab spots. On-chip activation of T-cells results in release of cytokines and detection of these secreted molecules on adjacent Ab spots. Sandwich immunoassay with colorimetric readout is used to visualize cytokine signal. ( C ) Ab microarrays are integrated with a microfluidic conduit to minimize blood volume required for analysis. T-cell capture and cytokine detection occurs in the microfluidic device.

    Journal: Analytical chemistry

    Article Title: Lensfree Holographic Imaging of Antibody Microarrays for High-Throughput Detection of Leukocyte Numbers and Function

    doi: 10.1021/ac100142a

    Figure Lengend Snippet: ( A ) Design of an antibody (Ab) microarray for capturing leukocytes and detecting secreted cytokines. T-cell Ab spots were print next to spots for detection of IFN-γ, TNF-α and IL-2. Anti-mouse IgG spots provided negative control whereas biotin spots were used for positive control. ( B ) Principle of operation of the assay. T-cells are captured in the immediate vicinity of cytokine-sensing Ab spots. On-chip activation of T-cells results in release of cytokines and detection of these secreted molecules on adjacent Ab spots. Sandwich immunoassay with colorimetric readout is used to visualize cytokine signal. ( C ) Ab microarrays are integrated with a microfluidic conduit to minimize blood volume required for analysis. T-cell capture and cytokine detection occurs in the microfluidic device.

    Article Snippet: Purified mouse anti-human IFN-γ Ab (clone K3.53), anti-IL2 Ab (clone 5355), anti-TNF-α/TNFSF1A Ab (clone 28401), biotinylated goat anti-human IFN-γ, IL2 and TNF-α/TNFSF1A Abs from R & D Systems (Minneapolis, MN).

    Techniques: Microarray, Negative Control, Positive Control, Chromatin Immunoprecipitation, Activation Assay

    Serum cytokine levels in PRRSV-infected pigs. Sera collected from pigs (n = 5) at 0, 3, 7, 14, 21 and 28 dpc were analysed for a IFN-α; b TNF-α; c IL-12; d IL-10; and e IL-4 by ELISA. Each bar represents the average cytokine amount from 5 pigs ± SEM. Bars showing different letters represent values that differ significantly from each other ( p

    Journal: BMC Veterinary Research

    Article Title: In vitro immune responses of porcine alveolar macrophages reflect host immune responses against porcine reproductive and respiratory syndrome viruses

    doi: 10.1186/s12917-018-1675-x

    Figure Lengend Snippet: Serum cytokine levels in PRRSV-infected pigs. Sera collected from pigs (n = 5) at 0, 3, 7, 14, 21 and 28 dpc were analysed for a IFN-α; b TNF-α; c IL-12; d IL-10; and e IL-4 by ELISA. Each bar represents the average cytokine amount from 5 pigs ± SEM. Bars showing different letters represent values that differ significantly from each other ( p

    Article Snippet: To detect IFN-α protein levels in sera and cell lysates, 100 μL (1.8 μg/mL) of a mouse anti-pig IFN-α antibody (Clone F17, PBL Assay Science, NJ, USA) was used as a coating antibody, and a mouse anti-pig IFN-α antibody (Clone K9, PBL Assay Science, NJ, USA) was biotinylated and used as a secondary antibody, with recombinant porcine IFN-α (PBL Assay Science, NJ, USA) as a standard.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay

    Evolution of IFN-α levels (Limit of detection: 3.9 pg/mL)

    Journal: Porcine Health Management

    Article Title: Immune response development after vaccination of 1-day-old naïve pigs with a Porcine Reproductive and Respiratory Syndrome 1-based modified live virus vaccine

    doi: 10.1186/s40813-018-0112-7

    Figure Lengend Snippet: Evolution of IFN-α levels (Limit of detection: 3.9 pg/mL)

    Article Snippet: Briefly, plates were coated with anti-pig IFN-α (mAb K9, R & D systems) at 1.1 Ug/mL; 50 μl of each sample and biotinylated pig IFN-α (mAb F17, R & D systems) were added.

    Techniques: