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human tnf-alpha duoset elisa  (Bio-Techne corporation)


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    Structured Review

    Bio-Techne corporation human tnf-alpha duoset elisa
    Human Tnf Alpha Duoset Elisa, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 98/100, based on 1202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+tnf-alpha+duoset+elisa/Human+TNF-alpha+DuoSet+ELISA/custom%40dy210%4042156999
    Average 98 stars, based on 1202 article reviews
    human tnf-alpha duoset elisa - by Bioz Stars, 2026-10
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    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Oligomer Restriction:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Generated:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    SYBR Green Assay:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    cDNA Synthesis:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Polymerase Chain Reaction:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Staining:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Isolation:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Cell Isolation:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Modification:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Recombinant:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Control:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Cell Culture:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (

    Blocking Assay:

    Article Title: Long-term immunoprophylaxis by TLR9 ligand mediates macrophage-ILC cross-talk against respiratory viral infection
    Article Snippet: a density of 1 × 10 5 cells per well and stimulated with K3-SPG (10 μg/ml) or zymosan depleted (Invivogen, 5 μg/ml). Supernatants were subjected to enzyme-linked immunosorbent assay (ELISA) for TNF-α (R&D Systems, #DY210) and IL-6 (R&D Systems, #DY206). Experiment was conducted with approval and according to the institutional guidelines of the Institute of Medical Science, The University of Tokyo (



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    Plant extract mixture downregulates inflammatory markers in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) IL1A , ( b ) IL1B, ( c ) IL1RN , ( d ) <t>CXCL8</t> , ( e ) TNFA , ( f ) IL17C , ( g ) IL36G , ( h ) CSF2 , ( i ) VEGFA were measured. Statistical significance was tested by a, b, c, e, h) one-way ANOVA with subsequent Sidak’s multiple comparison test or d, f, g) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). Protein expression levels of ( j ) IL-8 and <t>k)</t> <t>TNFα</t> were measured in the supernatants of the cells by <t>ELISA.</t> Values for unstimulated control and plant extract-treated cells were below the detection limit of 31,3 pg/mL (IL-8) or 62,5 pg/mL (TNFα) and not detectable; therefore, no statistical analysis was performed. n.d.= non-detectable.
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    ( a ) IL-6 and IL-8 from the basolateral sides of the ALI were determined using the <t>DuoSet®</t> <t>ELISA</t> kits. Data are presented as the mean ± SD (n = 4). Statistical p -values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the all treatments to the DMSO control; ¥, p < 0.05; *, p < 0.01. ( b ) Representative Western blot images showing CFTR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), and NF-κB p65 expression in treated polarized 16HBE14o-cells in ( a ). All data were obtained from three independent experiments.
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    Determination of M1 vs. M2 polarization in iMac. a Pro-inflammatory M1 cytokines IL-1β, <t>TNFα,</t> IL-6, and IL-8 measured <t>by</t> <t>ELISA.</t> b Anti-inflammatory M2 cytokines IL-4, IL-10, IL-13, CCL5, CCL17, and CCL20 measured by ELISA. For a , b iMac were cultured in 6-well plates (3 × 10 6 cells/well). To challenge with tumor factors, iMac were co-cultured with 50% fresh supernatant from SK-MEL-28 cells every 24 h for 3 days. Supernatants were collected during the final 24 h for cytokine measurement. c Activation (phosphorylation) of NF-kB p65, STAT1, STAT3, and STAT6 in iMac was determined using Western blotting after conditioning with melanoma (SK-MEL-28) supernatant for 72 h. Protein band intensity was quantified using Odyssey version 3.0 software. After adjusting for loading using the β-actin band, data are presented as fold change to the iMac (Dox-) group, which is set as 1. For a – c , one-way ANOVA with Bonferroni’s correction for multiple comparisons was used to analyze group comparisons. Data are presented as means ± SD. Each experiment was performed three times, with triplicate samples each time. * p < 0.05, *** p < 0.001, and **** p < 0.0001
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    Image Search Results


    Plant extract mixture downregulates inflammatory markers in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) IL1A , ( b ) IL1B, ( c ) IL1RN , ( d ) CXCL8 , ( e ) TNFA , ( f ) IL17C , ( g ) IL36G , ( h ) CSF2 , ( i ) VEGFA were measured. Statistical significance was tested by a, b, c, e, h) one-way ANOVA with subsequent Sidak’s multiple comparison test or d, f, g) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). Protein expression levels of ( j ) IL-8 and k) TNFα were measured in the supernatants of the cells by ELISA. Values for unstimulated control and plant extract-treated cells were below the detection limit of 31,3 pg/mL (IL-8) or 62,5 pg/mL (TNFα) and not detectable; therefore, no statistical analysis was performed. n.d.= non-detectable.

    Journal: Scientific Reports

    Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

    doi: 10.1038/s41598-026-50000-8

    Figure Lengend Snippet: Plant extract mixture downregulates inflammatory markers in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) IL1A , ( b ) IL1B, ( c ) IL1RN , ( d ) CXCL8 , ( e ) TNFA , ( f ) IL17C , ( g ) IL36G , ( h ) CSF2 , ( i ) VEGFA were measured. Statistical significance was tested by a, b, c, e, h) one-way ANOVA with subsequent Sidak’s multiple comparison test or d, f, g) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). Protein expression levels of ( j ) IL-8 and k) TNFα were measured in the supernatants of the cells by ELISA. Values for unstimulated control and plant extract-treated cells were below the detection limit of 31,3 pg/mL (IL-8) or 62,5 pg/mL (TNFα) and not detectable; therefore, no statistical analysis was performed. n.d.= non-detectable.

    Article Snippet: TNFα and IL-8 concentrations were determined using commercial ELISA kits (human TNFα or human CXCL8/IL-8 DuoSet ELISA, R&D Systems, Minneapolis, MN, USA) following the manufacturer’s protocol.

    Techniques: Plant Extract, Gene Expression, Comparison, Expressing, Enzyme-linked Immunosorbent Assay, Control

    ( a ) IL-6 and IL-8 from the basolateral sides of the ALI were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n = 4). Statistical p -values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the all treatments to the DMSO control; ¥, p < 0.05; *, p < 0.01. ( b ) Representative Western blot images showing CFTR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), and NF-κB p65 expression in treated polarized 16HBE14o-cells in ( a ). All data were obtained from three independent experiments.

    Journal: bioRxiv

    Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

    doi: 10.64898/2026.04.21.719931

    Figure Lengend Snippet: ( a ) IL-6 and IL-8 from the basolateral sides of the ALI were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n = 4). Statistical p -values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the all treatments to the DMSO control; ¥, p < 0.05; *, p < 0.01. ( b ) Representative Western blot images showing CFTR, phospho-AKT (Ser473), AKT, phospho-NF-κB p65 (Ser536), and NF-κB p65 expression in treated polarized 16HBE14o-cells in ( a ). All data were obtained from three independent experiments.

    Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

    Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing

    ALI cells were treated with DMSO, 100 ng/mL PM 2.5, 10 µM LY 294002 ± PM 2.5, or 10 µM LY 303511 ± PM 2.5 for 24 hrs. ( a ) IL-6 and IL-8 from the basolateral sides were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n=3). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing all treatments to the DMSO control, or PM 2.5 to PM 2.5 ± LY 294002 or LY 303511; *, p < 0.01. ( b ) Representative Western blot images showing CFTR and β-actin expression. β-actin was used to ensure equal loading of protein. All data were obtained from three independent experiments, and each experiment was analyzed in duplication.

    Journal: bioRxiv

    Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

    doi: 10.64898/2026.04.21.719931

    Figure Lengend Snippet: ALI cells were treated with DMSO, 100 ng/mL PM 2.5, 10 µM LY 294002 ± PM 2.5, or 10 µM LY 303511 ± PM 2.5 for 24 hrs. ( a ) IL-6 and IL-8 from the basolateral sides were determined using the DuoSet® ELISA kits. Data are presented as the mean ± SD (n=3). Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing all treatments to the DMSO control, or PM 2.5 to PM 2.5 ± LY 294002 or LY 303511; *, p < 0.01. ( b ) Representative Western blot images showing CFTR and β-actin expression. β-actin was used to ensure equal loading of protein. All data were obtained from three independent experiments, and each experiment was analyzed in duplication.

    Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

    Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Expressing

    ( a ) The lung parenchyma from two ferrets were sliced into several sections as shown. Arrows indicate the tracheobronchial tubes. All tissue sections were incubating with DMSO (control), 10 µM B[a]P, or 100 ng/mL PM 2.5 in Pneumacult-EX Plus Basal media for 24 hrs. ( b ) IL 6 secretion was determined using the ferret ELISA kit. Data represent the mean of 6 independent sections. Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; ¥, p < 0.05 (n = 6). ( c ) Representative western blotting images show reduction of CFTR, and β2AR and elevation of TRPC6 expression in explants under these treatments.

    Journal: bioRxiv

    Article Title: PM2.5 toxin benzo[a]pyrene induces life-limiting inflammation and oxidative stress in the airway by up-regulation of TRPC6 and inactivation of β2AR/CFTR signaling

    doi: 10.64898/2026.04.21.719931

    Figure Lengend Snippet: ( a ) The lung parenchyma from two ferrets were sliced into several sections as shown. Arrows indicate the tracheobronchial tubes. All tissue sections were incubating with DMSO (control), 10 µM B[a]P, or 100 ng/mL PM 2.5 in Pneumacult-EX Plus Basal media for 24 hrs. ( b ) IL 6 secretion was determined using the ferret ELISA kit. Data represent the mean of 6 independent sections. Statistical p values were determined with a one-way ANOVA, followed by the Tukey post-hoc test comparing the B[a]P or PM 2.5 treatments to the DMSO control; ¥, p < 0.05 (n = 6). ( c ) Representative western blotting images show reduction of CFTR, and β2AR and elevation of TRPC6 expression in explants under these treatments.

    Article Snippet: Benzo[a]pyrene, PM 2.5 (ERMCZ110), CFTR inh -172, IBMX, amiloride, Forskolin, MG-132, L-glutathione, and Millipore Immobilon Western (Sigma-Aldrich); BI 749327, LY 294002, and LY 303511 (Cayman Chemical); MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide; EZ-LinkTM Sulfo-NHS-SS-Biotin, CellROX green reagent, Dynabeads Protein G, Pierce Streptavidin magnetic beads, HaltTM protease and Phosphatase Inhibitor Cocktail (Thermo Fisher) ; Fluo-4 AM and Probenecid (Invitrogen); Human IL-8/CXCL8 DuoSet® ELISA kit, Human IL-6 ELISA DuoSet® kit, and Human TNF-alpha DuoSet® ELISA (R&D Systems); Ferret IL-6 ELISA kit (Genorise, PA, USA); Pneumacult-ALI media kit and Pneumacult-EX Plus Basal media kit (Stemcell); and Gibco MEM (Thermo Fisher).

    Techniques: Control, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

    Determination of M1 vs. M2 polarization in iMac. a Pro-inflammatory M1 cytokines IL-1β, TNFα, IL-6, and IL-8 measured by ELISA. b Anti-inflammatory M2 cytokines IL-4, IL-10, IL-13, CCL5, CCL17, and CCL20 measured by ELISA. For a , b iMac were cultured in 6-well plates (3 × 10 6 cells/well). To challenge with tumor factors, iMac were co-cultured with 50% fresh supernatant from SK-MEL-28 cells every 24 h for 3 days. Supernatants were collected during the final 24 h for cytokine measurement. c Activation (phosphorylation) of NF-kB p65, STAT1, STAT3, and STAT6 in iMac was determined using Western blotting after conditioning with melanoma (SK-MEL-28) supernatant for 72 h. Protein band intensity was quantified using Odyssey version 3.0 software. After adjusting for loading using the β-actin band, data are presented as fold change to the iMac (Dox-) group, which is set as 1. For a – c , one-way ANOVA with Bonferroni’s correction for multiple comparisons was used to analyze group comparisons. Data are presented as means ± SD. Each experiment was performed three times, with triplicate samples each time. * p < 0.05, *** p < 0.001, and **** p < 0.0001

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Bioengineered iPSC-derived human macrophages with increased angiotensin-converting enzyme (ACE) expression suppress solid tumor growth

    doi: 10.1038/s41392-026-02650-3

    Figure Lengend Snippet: Determination of M1 vs. M2 polarization in iMac. a Pro-inflammatory M1 cytokines IL-1β, TNFα, IL-6, and IL-8 measured by ELISA. b Anti-inflammatory M2 cytokines IL-4, IL-10, IL-13, CCL5, CCL17, and CCL20 measured by ELISA. For a , b iMac were cultured in 6-well plates (3 × 10 6 cells/well). To challenge with tumor factors, iMac were co-cultured with 50% fresh supernatant from SK-MEL-28 cells every 24 h for 3 days. Supernatants were collected during the final 24 h for cytokine measurement. c Activation (phosphorylation) of NF-kB p65, STAT1, STAT3, and STAT6 in iMac was determined using Western blotting after conditioning with melanoma (SK-MEL-28) supernatant for 72 h. Protein band intensity was quantified using Odyssey version 3.0 software. After adjusting for loading using the β-actin band, data are presented as fold change to the iMac (Dox-) group, which is set as 1. For a – c , one-way ANOVA with Bonferroni’s correction for multiple comparisons was used to analyze group comparisons. Data are presented as means ± SD. Each experiment was performed three times, with triplicate samples each time. * p < 0.05, *** p < 0.001, and **** p < 0.0001

    Article Snippet: We used a commercial solid-phase ELISA kit protocol to measure IL-1β (Invitrogen, BMS224-2), TNFα (R&D Systems, DY210-05), IL-6 (R&D Systems, DY206-05), IL-8 (R&D Systems, DY208-05), IL-4 (R&D Systems, DY204-05), IL-10 (R&D Systems, DY217B-05), IL-13 (R&D Systems, DY213-05), CCL5 (R&D Systems, DY278-05), CCL17 (R&D Systems, DY364-05), and CCL20 (R&D Systems, DY360-05).

    Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Activation Assay, Phospho-proteomics, Western Blot, Software