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BioLegend plasma il 6 levels
NK cell depletion reduces lung viral burden and lung IFN-γ in vaccinated, influenza-challenged mice. Four days post infection in the model described in Fig. 2A , (A) Lung RNA was analyzed by qPCR for influenza viral burden (plotted against a dose curve of IFA with known HAU, giving HAU equivalents per 5ug RNA tested). (B-C) Transcript levels of inflammatory cytokine genes (B) Il6 and Ifnγ and (C) neutrophil-related chemokines Cxcl1 and Cxcl2 , along with neutrophil lipocalin protein (Lcn2) RNA induction normalized to housekeeping gene ß-actin and displayed as induction over mock-treated control mice. (D) Plasma levels of <t>IL-6</t> (pg/mL). (E) IL-6 levels in lung supernatants after whole lung enzymatic digestion for single cell isolation and viral burden quantification. (A-E) A pool of two independent experiments (n=5-9/group; (A-D) were female {F} and (E) were male {E} mice used for single cell isolation). Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney U test, ns = not significant. Spearman correlation coefficient shown.
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Article Title: Natural Killer cells dampen the pathogenic features of recall responses to influenza infection

Journal: bioRxiv

doi: 10.1101/846626

NK cell depletion reduces lung viral burden and lung IFN-γ in vaccinated, influenza-challenged mice. Four days post infection in the model described in Fig. 2A , (A) Lung RNA was analyzed by qPCR for influenza viral burden (plotted against a dose curve of IFA with known HAU, giving HAU equivalents per 5ug RNA tested). (B-C) Transcript levels of inflammatory cytokine genes (B) Il6 and Ifnγ and (C) neutrophil-related chemokines Cxcl1 and Cxcl2 , along with neutrophil lipocalin protein (Lcn2) RNA induction normalized to housekeeping gene ß-actin and displayed as induction over mock-treated control mice. (D) Plasma levels of IL-6 (pg/mL). (E) IL-6 levels in lung supernatants after whole lung enzymatic digestion for single cell isolation and viral burden quantification. (A-E) A pool of two independent experiments (n=5-9/group; (A-D) were female {F} and (E) were male {E} mice used for single cell isolation). Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney U test, ns = not significant. Spearman correlation coefficient shown.
Figure Legend Snippet: NK cell depletion reduces lung viral burden and lung IFN-γ in vaccinated, influenza-challenged mice. Four days post infection in the model described in Fig. 2A , (A) Lung RNA was analyzed by qPCR for influenza viral burden (plotted against a dose curve of IFA with known HAU, giving HAU equivalents per 5ug RNA tested). (B-C) Transcript levels of inflammatory cytokine genes (B) Il6 and Ifnγ and (C) neutrophil-related chemokines Cxcl1 and Cxcl2 , along with neutrophil lipocalin protein (Lcn2) RNA induction normalized to housekeeping gene ß-actin and displayed as induction over mock-treated control mice. (D) Plasma levels of IL-6 (pg/mL). (E) IL-6 levels in lung supernatants after whole lung enzymatic digestion for single cell isolation and viral burden quantification. (A-E) A pool of two independent experiments (n=5-9/group; (A-D) were female {F} and (E) were male {E} mice used for single cell isolation). Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney U test, ns = not significant. Spearman correlation coefficient shown.

Techniques Used: Mouse Assay, Infection, Real-time Polymerase Chain Reaction, Immunofluorescence, Single-cell Isolation, MANN-WHITNEY

Depletion of NK cells after vaccination, and subsequent repopulation, does not alter lung viral burden, disease severity or systemic inflammation after challenge. (A) Transgenic C57BL/6 mice with NKp46 driven expression of diphtheria toxin (DT) receptor were vaccinated 42 days (d) prior to intranasal influenza (Flu) challenge and treated with DT (NK-depleted) 21 days prior to challenge with necropsy (nx) at 4 days post influenza challenge. (B) At 3 and 21 days post DT treatment, lungs were excised and single cells isolated for flow cytometry for the proportion (%) of NK1.1+, NKp46+ NK cells. (C) Weight loss at 4 days post influenza challenge. (D) Lung cell-free supernatants were analyzed by qPCR for influenza viral burden (plotted against a dose curve of Flu with known HAU, giving HAU equivalents). (E) Plasma levels of IL-6 (pg/mL). (B) NK cell depletion data a pool of two independent experiments, (n=2/group of male {M} mice) and (n=5-6/group of female {F} mice). (C-D) Data a pool of two independent experiment (n=4-10/group); all male {M} mice. Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney test, ns = not significant.
Figure Legend Snippet: Depletion of NK cells after vaccination, and subsequent repopulation, does not alter lung viral burden, disease severity or systemic inflammation after challenge. (A) Transgenic C57BL/6 mice with NKp46 driven expression of diphtheria toxin (DT) receptor were vaccinated 42 days (d) prior to intranasal influenza (Flu) challenge and treated with DT (NK-depleted) 21 days prior to challenge with necropsy (nx) at 4 days post influenza challenge. (B) At 3 and 21 days post DT treatment, lungs were excised and single cells isolated for flow cytometry for the proportion (%) of NK1.1+, NKp46+ NK cells. (C) Weight loss at 4 days post influenza challenge. (D) Lung cell-free supernatants were analyzed by qPCR for influenza viral burden (plotted against a dose curve of Flu with known HAU, giving HAU equivalents). (E) Plasma levels of IL-6 (pg/mL). (B) NK cell depletion data a pool of two independent experiments, (n=2/group of male {M} mice) and (n=5-6/group of female {F} mice). (C-D) Data a pool of two independent experiment (n=4-10/group); all male {M} mice. Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney test, ns = not significant.

Techniques Used: Transgenic Assay, Mouse Assay, Expressing, Isolation, Flow Cytometry, Real-time Polymerase Chain Reaction, MANN-WHITNEY

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Enzyme-linked Immunosorbent Assay:

Article Title: Natural Killer cells dampen the pathogenic features of recall responses to influenza infection
Article Snippet: .. Plasma IL-6 levels were determined by sandwich enzyme-linked immunosorbent assay (ELISA) (Biolegend ELISA MAX Deluxe, London UK). .. To determine circulating influenza HA antibodies following influenza vaccination, a direct ELISA was done.

Sandwich ELISA:

Article Title: Natural Killer cells dampen the pathogenic features of recall responses to influenza infection
Article Snippet: .. Plasma IL-6 levels were determined by sandwich enzyme-linked immunosorbent assay (ELISA) (Biolegend ELISA MAX Deluxe, London UK). .. To determine circulating influenza HA antibodies following influenza vaccination, a direct ELISA was done.

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    BioLegend plasma il 6 levels
    NK cell depletion reduces lung viral burden and lung IFN-γ in vaccinated, influenza-challenged mice. Four days post infection in the model described in Fig. 2A , (A) Lung RNA was analyzed by qPCR for influenza viral burden (plotted against a dose curve of IFA with known HAU, giving HAU equivalents per 5ug RNA tested). (B-C) Transcript levels of inflammatory cytokine genes (B) Il6 and Ifnγ and (C) neutrophil-related chemokines Cxcl1 and Cxcl2 , along with neutrophil lipocalin protein (Lcn2) RNA induction normalized to housekeeping gene ß-actin and displayed as induction over mock-treated control mice. (D) Plasma levels of <t>IL-6</t> (pg/mL). (E) IL-6 levels in lung supernatants after whole lung enzymatic digestion for single cell isolation and viral burden quantification. (A-E) A pool of two independent experiments (n=5-9/group; (A-D) were female {F} and (E) were male {E} mice used for single cell isolation). Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney U test, ns = not significant. Spearman correlation coefficient shown.
    Plasma Il 6 Levels, supplied by BioLegend, used in various techniques. Bioz Stars score: 89/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend il 21
    Relationship between plasma cytokines and total IgE in patients with chronic spontaneous urticaria (CSU). A. Concentrations of IL-4, IL-6, <t>IL-21</t> and total IgE in the plasma of CSU patients and healthy controls (HC). Differences between CSU and HC were evaluated with a two-tailed Student’s t -test. B. Correlation analysis between IL-4, IL-6, IL-21 and total IgE were conducted using Spearman’s rank test. Each plot represented one sample. P
    Il 21, supplied by BioLegend, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend hyaluronan
    Effects of water immersion stress treatment on the plasma levels of ROS, IL-6, corticosterone, and <t>hyaluronan</t> in arthritic mice. Plasma levels of these proteins were assessed by enzyme-linked immunosorbent assay. Data are presented as means ± standard deviations (n = 5). * P
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    NK cell depletion reduces lung viral burden and lung IFN-γ in vaccinated, influenza-challenged mice. Four days post infection in the model described in Fig. 2A , (A) Lung RNA was analyzed by qPCR for influenza viral burden (plotted against a dose curve of IFA with known HAU, giving HAU equivalents per 5ug RNA tested). (B-C) Transcript levels of inflammatory cytokine genes (B) Il6 and Ifnγ and (C) neutrophil-related chemokines Cxcl1 and Cxcl2 , along with neutrophil lipocalin protein (Lcn2) RNA induction normalized to housekeeping gene ß-actin and displayed as induction over mock-treated control mice. (D) Plasma levels of IL-6 (pg/mL). (E) IL-6 levels in lung supernatants after whole lung enzymatic digestion for single cell isolation and viral burden quantification. (A-E) A pool of two independent experiments (n=5-9/group; (A-D) were female {F} and (E) were male {E} mice used for single cell isolation). Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney U test, ns = not significant. Spearman correlation coefficient shown.

    Journal: bioRxiv

    Article Title: Natural Killer cells dampen the pathogenic features of recall responses to influenza infection

    doi: 10.1101/846626

    Figure Lengend Snippet: NK cell depletion reduces lung viral burden and lung IFN-γ in vaccinated, influenza-challenged mice. Four days post infection in the model described in Fig. 2A , (A) Lung RNA was analyzed by qPCR for influenza viral burden (plotted against a dose curve of IFA with known HAU, giving HAU equivalents per 5ug RNA tested). (B-C) Transcript levels of inflammatory cytokine genes (B) Il6 and Ifnγ and (C) neutrophil-related chemokines Cxcl1 and Cxcl2 , along with neutrophil lipocalin protein (Lcn2) RNA induction normalized to housekeeping gene ß-actin and displayed as induction over mock-treated control mice. (D) Plasma levels of IL-6 (pg/mL). (E) IL-6 levels in lung supernatants after whole lung enzymatic digestion for single cell isolation and viral burden quantification. (A-E) A pool of two independent experiments (n=5-9/group; (A-D) were female {F} and (E) were male {E} mice used for single cell isolation). Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney U test, ns = not significant. Spearman correlation coefficient shown.

    Article Snippet: Plasma IL-6 levels were determined by sandwich enzyme-linked immunosorbent assay (ELISA) (Biolegend ELISA MAX Deluxe, London UK).

    Techniques: Mouse Assay, Infection, Real-time Polymerase Chain Reaction, Immunofluorescence, Single-cell Isolation, MANN-WHITNEY

    Depletion of NK cells after vaccination, and subsequent repopulation, does not alter lung viral burden, disease severity or systemic inflammation after challenge. (A) Transgenic C57BL/6 mice with NKp46 driven expression of diphtheria toxin (DT) receptor were vaccinated 42 days (d) prior to intranasal influenza (Flu) challenge and treated with DT (NK-depleted) 21 days prior to challenge with necropsy (nx) at 4 days post influenza challenge. (B) At 3 and 21 days post DT treatment, lungs were excised and single cells isolated for flow cytometry for the proportion (%) of NK1.1+, NKp46+ NK cells. (C) Weight loss at 4 days post influenza challenge. (D) Lung cell-free supernatants were analyzed by qPCR for influenza viral burden (plotted against a dose curve of Flu with known HAU, giving HAU equivalents). (E) Plasma levels of IL-6 (pg/mL). (B) NK cell depletion data a pool of two independent experiments, (n=2/group of male {M} mice) and (n=5-6/group of female {F} mice). (C-D) Data a pool of two independent experiment (n=4-10/group); all male {M} mice. Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney test, ns = not significant.

    Journal: bioRxiv

    Article Title: Natural Killer cells dampen the pathogenic features of recall responses to influenza infection

    doi: 10.1101/846626

    Figure Lengend Snippet: Depletion of NK cells after vaccination, and subsequent repopulation, does not alter lung viral burden, disease severity or systemic inflammation after challenge. (A) Transgenic C57BL/6 mice with NKp46 driven expression of diphtheria toxin (DT) receptor were vaccinated 42 days (d) prior to intranasal influenza (Flu) challenge and treated with DT (NK-depleted) 21 days prior to challenge with necropsy (nx) at 4 days post influenza challenge. (B) At 3 and 21 days post DT treatment, lungs were excised and single cells isolated for flow cytometry for the proportion (%) of NK1.1+, NKp46+ NK cells. (C) Weight loss at 4 days post influenza challenge. (D) Lung cell-free supernatants were analyzed by qPCR for influenza viral burden (plotted against a dose curve of Flu with known HAU, giving HAU equivalents). (E) Plasma levels of IL-6 (pg/mL). (B) NK cell depletion data a pool of two independent experiments, (n=2/group of male {M} mice) and (n=5-6/group of female {F} mice). (C-D) Data a pool of two independent experiment (n=4-10/group); all male {M} mice. Dots represent individual mice with bars showing mean. Significance determined by Mann-Whitney test, ns = not significant.

    Article Snippet: Plasma IL-6 levels were determined by sandwich enzyme-linked immunosorbent assay (ELISA) (Biolegend ELISA MAX Deluxe, London UK).

    Techniques: Transgenic Assay, Mouse Assay, Expressing, Isolation, Flow Cytometry, Real-time Polymerase Chain Reaction, MANN-WHITNEY

    Relationship between plasma cytokines and total IgE in patients with chronic spontaneous urticaria (CSU). A. Concentrations of IL-4, IL-6, IL-21 and total IgE in the plasma of CSU patients and healthy controls (HC). Differences between CSU and HC were evaluated with a two-tailed Student’s t -test. B. Correlation analysis between IL-4, IL-6, IL-21 and total IgE were conducted using Spearman’s rank test. Each plot represented one sample. P

    Journal: American Journal of Translational Research

    Article Title: Pathogenic role of circulating CD4+CXCR5+ cell subpopulations in patients with chronic spontaneous urticarial

    doi:

    Figure Lengend Snippet: Relationship between plasma cytokines and total IgE in patients with chronic spontaneous urticaria (CSU). A. Concentrations of IL-4, IL-6, IL-21 and total IgE in the plasma of CSU patients and healthy controls (HC). Differences between CSU and HC were evaluated with a two-tailed Student’s t -test. B. Correlation analysis between IL-4, IL-6, IL-21 and total IgE were conducted using Spearman’s rank test. Each plot represented one sample. P

    Article Snippet: Correlation between the plasma levels of IL-4, IL-6, IL-21 and the total IgE in CSUSignificantly higher plasma levels of IL-4 and IL-6 but lower IL-21 were found in patients with CSU than in HC ( ).

    Techniques: Two Tailed Test

    Effects of water immersion stress treatment on the plasma levels of ROS, IL-6, corticosterone, and hyaluronan in arthritic mice. Plasma levels of these proteins were assessed by enzyme-linked immunosorbent assay. Data are presented as means ± standard deviations (n = 5). * P

    Journal: bioRxiv

    Article Title: Deterioration of dry skin in arthritis model mice via stress-induced changes in immune cells in the thymus and spleen

    doi: 10.1101/641720

    Figure Lengend Snippet: Effects of water immersion stress treatment on the plasma levels of ROS, IL-6, corticosterone, and hyaluronan in arthritic mice. Plasma levels of these proteins were assessed by enzyme-linked immunosorbent assay. Data are presented as means ± standard deviations (n = 5). * P

    Article Snippet: Plasma levels of IL-6, corticosterone, and hyaluronan were measured using commercial enzyme-linked immunosorbent assay kits (IL-6: BioLegend, San Diego, CA, USA; corticosterone: Enzo Life Science Inc., Farmingdale, NY, USA; hyaluronan: R & D Systems, Minneapolis, MN, USA), according to the manufacturers’ instructions.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay