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Protein levels and relative gene expression levels of IL-8 after mitogen stimulation of PBMCs. (a) IL-8 median fluorescent intensity (MFI) in supernatants from unstimulated and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using <t>MILLIPLEX®</t> Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median MFI in supernatants. Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Significantly different from unstimulated samples: * P = 0.05. ns: Not significantly different from unstimulated samples (P < 0.05). The kit-specific standards used to generate a 7-point standard curve often deviated from the recovery range, results are therefore reported as MFI and not converted to concentration (pg/mL). (b) Relative gene expression levels after mitogen stimulation of PBMC. Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference
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Protein levels and relative gene expression levels of IL-8 after mitogen stimulation of PBMCs. (a) IL-8 median fluorescent intensity (MFI) in supernatants from unstimulated and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using <t>MILLIPLEX®</t> Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median MFI in supernatants. Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Significantly different from unstimulated samples: * P = 0.05. ns: Not significantly different from unstimulated samples (P < 0.05). The kit-specific standards used to generate a 7-point standard curve often deviated from the recovery range, results are therefore reported as MFI and not converted to concentration (pg/mL). (b) Relative gene expression levels after mitogen stimulation of PBMC. Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference
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Protein levels and relative gene expression levels of IL-8 after mitogen stimulation of PBMCs. (a) IL-8 median fluorescent intensity (MFI) in supernatants from unstimulated and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using <t>MILLIPLEX®</t> Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median MFI in supernatants. Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Significantly different from unstimulated samples: * P = 0.05. ns: Not significantly different from unstimulated samples (P < 0.05). The kit-specific standards used to generate a 7-point standard curve often deviated from the recovery range, results are therefore reported as MFI and not converted to concentration (pg/mL). (b) Relative gene expression levels after mitogen stimulation of PBMC. Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference
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LINCO bead-based assay human cytokine/chemokine multiplex immunoassay kits
Protein levels and relative gene expression levels of IL-8 after mitogen stimulation of PBMCs. (a) IL-8 median fluorescent intensity (MFI) in supernatants from unstimulated and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using <t>MILLIPLEX®</t> Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median MFI in supernatants. Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Significantly different from unstimulated samples: * P = 0.05. ns: Not significantly different from unstimulated samples (P < 0.05). The kit-specific standards used to generate a 7-point standard curve often deviated from the recovery range, results are therefore reported as MFI and not converted to concentration (pg/mL). (b) Relative gene expression levels after mitogen stimulation of PBMC. Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference
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Protein levels and relative gene expression levels of IL-8 after mitogen stimulation of PBMCs. (a) IL-8 median fluorescent intensity (MFI) in supernatants from unstimulated and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using MILLIPLEX® Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median MFI in supernatants. Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Significantly different from unstimulated samples: * P = 0.05. ns: Not significantly different from unstimulated samples (P < 0.05). The kit-specific standards used to generate a 7-point standard curve often deviated from the recovery range, results are therefore reported as MFI and not converted to concentration (pg/mL). (b) Relative gene expression levels after mitogen stimulation of PBMC. Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference

Journal: Acta Veterinaria Scandinavica

Article Title: Measuring cytokines in Eurasian tundra reindeer ( Rangifer tarandus tarandus ) with a bovine bead-based multiplex immunoassay and real-time PCR

doi: 10.1186/s13028-025-00819-4

Figure Lengend Snippet: Protein levels and relative gene expression levels of IL-8 after mitogen stimulation of PBMCs. (a) IL-8 median fluorescent intensity (MFI) in supernatants from unstimulated and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using MILLIPLEX® Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median MFI in supernatants. Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Significantly different from unstimulated samples: * P = 0.05. ns: Not significantly different from unstimulated samples (P < 0.05). The kit-specific standards used to generate a 7-point standard curve often deviated from the recovery range, results are therefore reported as MFI and not converted to concentration (pg/mL). (b) Relative gene expression levels after mitogen stimulation of PBMC. Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference

Article Snippet: Cytokine levels for IL-6, IL-8, IL-10, IL-17, TNF-α, and IFN-γ were measured using a bovine bead-based multiplex immunoassay (MILLIPLEX ® Bovine Cytokine/Chemokine Magnetic Bead multiplex assay kit, Merck Millipore, Darmstadt, Germany).

Techniques: Gene Expression, Multiplex Assay, Control, Incubation, Concentration Assay, Standard Deviation

IL-6 concentration after mitogen stimulation of PBMCs. IL-6 concentrations in supernatants from unstimulated controls and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs) measured using MILLIPLEX® Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicates median protein concentration (pg/mL) Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Open symbols (○, ∆) indicate points higher or lower than the limits of quantification (LOQ). Not significantly (ns) different from unstimulated samples (P < 0.05)

Journal: Acta Veterinaria Scandinavica

Article Title: Measuring cytokines in Eurasian tundra reindeer ( Rangifer tarandus tarandus ) with a bovine bead-based multiplex immunoassay and real-time PCR

doi: 10.1186/s13028-025-00819-4

Figure Lengend Snippet: IL-6 concentration after mitogen stimulation of PBMCs. IL-6 concentrations in supernatants from unstimulated controls and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs) measured using MILLIPLEX® Bovine Cytokine/Chemokine multiplex assay. The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicates median protein concentration (pg/mL) Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Open symbols (○, ∆) indicate points higher or lower than the limits of quantification (LOQ). Not significantly (ns) different from unstimulated samples (P < 0.05)

Article Snippet: Cytokine levels for IL-6, IL-8, IL-10, IL-17, TNF-α, and IFN-γ were measured using a bovine bead-based multiplex immunoassay (MILLIPLEX ® Bovine Cytokine/Chemokine Magnetic Bead multiplex assay kit, Merck Millipore, Darmstadt, Germany).

Techniques: Concentration Assay, Multiplex Assay, Protein Concentration, Control, Incubation

Cytokine levels measured in serum samples from reindeer inoculated with ORFV or CvHV2. Quality control samples showed concentrations lower than the expected table values provided by the manufacturer, and results are therefore reported as median fluorescent intensity (MFI) and not converted to concentration (pg/mL). (a) Cytokine levels measured in serum samples from reindeer (n = 6) 0, 5, and 12 days post inoculation (p.i.) with Orf virus (ORFV) using the MILLIPLEX ® Bovine Cytokine/Chemokine multiplex assay. The different days of sampling are indicated for each cytokine. (b) Cytokine levels measured in serum samples from reindeer (n = 5) 0, 4, 7, and 10 days p.i. with Varicellovirus cervidalpha2 (CvHV2) using the MILLIPLEX® Bovine Cytokine/Chemokine multiplex assay. Reindeer B1 is a non-inoculated control animal. The different days of sampling are indicated for each cytokine. Significantly different from unstimulated samples: *** P < 0.001, ** P < 0.01, * P = 0.01

Journal: Acta Veterinaria Scandinavica

Article Title: Measuring cytokines in Eurasian tundra reindeer ( Rangifer tarandus tarandus ) with a bovine bead-based multiplex immunoassay and real-time PCR

doi: 10.1186/s13028-025-00819-4

Figure Lengend Snippet: Cytokine levels measured in serum samples from reindeer inoculated with ORFV or CvHV2. Quality control samples showed concentrations lower than the expected table values provided by the manufacturer, and results are therefore reported as median fluorescent intensity (MFI) and not converted to concentration (pg/mL). (a) Cytokine levels measured in serum samples from reindeer (n = 6) 0, 5, and 12 days post inoculation (p.i.) with Orf virus (ORFV) using the MILLIPLEX ® Bovine Cytokine/Chemokine multiplex assay. The different days of sampling are indicated for each cytokine. (b) Cytokine levels measured in serum samples from reindeer (n = 5) 0, 4, 7, and 10 days p.i. with Varicellovirus cervidalpha2 (CvHV2) using the MILLIPLEX® Bovine Cytokine/Chemokine multiplex assay. Reindeer B1 is a non-inoculated control animal. The different days of sampling are indicated for each cytokine. Significantly different from unstimulated samples: *** P < 0.001, ** P < 0.01, * P = 0.01

Article Snippet: Cytokine levels for IL-6, IL-8, IL-10, IL-17, TNF-α, and IFN-γ were measured using a bovine bead-based multiplex immunoassay (MILLIPLEX ® Bovine Cytokine/Chemokine Magnetic Bead multiplex assay kit, Merck Millipore, Darmstadt, Germany).

Techniques: Control, Concentration Assay, Virus, Multiplex Assay, Sampling

Protein concentrations and relative gene expression levels of cytokines after mitogen stimulation of PBMCs. (a-d) Protein concentrations in supernatants from unstimulated control and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using MILLIPLEX ® Bovine Cytokine/Chemokine multiplex assay . The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median protein concentration (pg/mL). Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Open symbols (○, ∆) indicate points higher or lower than the limits of quantification (LOQ). (e–h) Relative gene expression levels after mitogen stimulation of PBMC. The x-axis represents stimulation time in hours (h) for reindeer PBMCs (n = 3–4). Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference. Significantly different from unstimulated samples: *** P < 0.001, ** P < 0.01

Journal: Acta Veterinaria Scandinavica

Article Title: Measuring cytokines in Eurasian tundra reindeer ( Rangifer tarandus tarandus ) with a bovine bead-based multiplex immunoassay and real-time PCR

doi: 10.1186/s13028-025-00819-4

Figure Lengend Snippet: Protein concentrations and relative gene expression levels of cytokines after mitogen stimulation of PBMCs. (a-d) Protein concentrations in supernatants from unstimulated control and phorbol myristate acetate and ionomycin (PMA-I) stimulated peripheral blood mononuclear cells (PBMCs), measured using MILLIPLEX ® Bovine Cytokine/Chemokine multiplex assay . The x-axis represents stimulation time in hours (h), with reindeer data (n = 4) on the left and cattle data (n = 3) on the right. Vertical lines indicate median protein concentration (pg/mL). Points denote individual measurements for unstimulated control (●) and PMA-I incubated (▲) samples. Open symbols (○, ∆) indicate points higher or lower than the limits of quantification (LOQ). (e–h) Relative gene expression levels after mitogen stimulation of PBMC. The x-axis represents stimulation time in hours (h) for reindeer PBMCs (n = 3–4). Bars depict averages ± standard deviation (SD) of relative mRNA levels, expressed as fold change. Points denote the fold change for individual animals (▲). Calculations utilize the 2 −ΔΔCq method, with β2M in unstimulated PBMCs as a reference. Significantly different from unstimulated samples: *** P < 0.001, ** P < 0.01

Article Snippet: Cytokine levels for IL-6, IL-8, IL-10, IL-17, TNF-α, and IFN-γ were measured using a bovine bead-based multiplex immunoassay (MILLIPLEX ® Bovine Cytokine/Chemokine Magnetic Bead multiplex assay kit, Merck Millipore, Darmstadt, Germany).

Techniques: Gene Expression, Control, Multiplex Assay, Protein Concentration, Incubation, Standard Deviation