Protein levels of the cytokines CXCL‐10 and IL‐6 in the cell culture supernatants were determined using the following ELISA kits: CXCL‐10 and IL‐6 (R&D System) according to manufacturer's instructions. IFN‐beta in the cell culture supernatants was determined using the mouse IFN‐beta DuoSet ELISA kit (R&D System) according to manufacturer's instructions. IFNγ in the T‐EVs were determined using the Human IFNγ DuoSet ELISA kit (R&D System) according to manufacturer's instruction. Prior to analysis, the T‐EVs were lyzed in v/v 2.5% Triton‐X‐100, 0.05% Tween‐20.
Human ifn γ duoset elisa kit
Manufactured by R&D Systems
The Human IFN-γ DuoSet ELISA kit is a sandwich enzyme-linked immunosorbent assay (ELISA) for the quantitative measurement of human interferon gamma (IFN-γ) in cell culture supernates, serum, and plasma.
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Lab products found in correlation
Cxcl 10 and il 6, by R&D Systems (1 mentions)
Mouse ifn beta duoset elisa kit, by R&D Systems (1 mentions)
Elisa kits for il 6, by BioLegend (1 mentions)
Human il 12 p70 duoset elisa kit, by R&D Systems (1 mentions)
Human vegf duoset elisa kit, by R&D Systems (1 mentions)
Human tgf beta 1 duoset elisa kit, by R&D Systems (1 mentions)
Human il 10 duoset elisa kit, by R&D Systems (1 mentions)
Lymphocyte separation medium 1077, by PromoCell (1 mentions)
Human il 2 duoset elisa kit, by R&D Systems (1 mentions)
Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
5 protocols using human ifn γ duoset elisa kit
1
Cytokine Quantification in T-Cell Derived Extracellular Vesicles
2
Cytokine Quantification by ELISA
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Protein levels of IL-6, IL-12 p70, or IFN-γ in supernatants were evaluated using ELISA kits for IL-6 (Biolegend), IL-12 (Human IL-12 p70 DuoSet ELISA kit, R&D Systems), and IFN-γ (Human IFN-γ DuoSet ELISA kit, R&D Systems), following the manufacturer's instructions.
3
Quantification of Cytokine Secretion
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The cell culture supernatants were collected for cytokines detection. The concentrations of IFN-γ, TGF-β, VEGFA and IL-10 were measured by using the Human IFN-γ DuoSet ELISA kit (R&D system), Human TGF-beta 1 DuoSet ELISA kit (R&D system), Human VEGF DuoSet ELISA kit (R&D) and Human IL-10 DuoSet ELISA kit (R&D systme) respectively, following the manufacturer’s instructions.
4
Isolation and Culture of Human PBMCs
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Antibodies were purchased from BioLegend (PerCP/Cy5.5 anti-human CD3, 317336; PE anti-human CCR7, 353204; APC anti-human CD45RA; 304112), Miltenyi Biotec (anti-human CD4-VioBlue, 130-097-333; anti-human CD8-PE/Vio770, 130-096-556) or Dianova (goat IgG anti-human IgG (Fc)-RPE, 109-115-098). Human IFN-γ DuoSet ELISA kit (DY285) and Human IL-2 DuoSet ELISA kit (DY202) were obtained from R&D Systems. CellTrace CFSE Cell Proliferation Kit (C34554) was purchased from Thermo Fisher Scientific. FaDu, SKBR-3 and MCF-7 cells were obtained from different sources and cultured as described previously.34 (link) LIM1215 were obtained from Merck KGaA (10092301-1VL) and cultured in RPMI-1640 (Thermo Fisher Scientific, 11875), 10% fetal bovine serum (FBS) (Pan Biotech, P30-3309). Density gradient centrifugation (Lymphocyte Separation Medium 1077, PromoCell, C-44010) was used to isolate human PBMCs from buffy coats of healthy donors (Blood Bank, Klinikum Stuttgart). PBMCs were cultivated in RPMI-1640, 10% FBS.
5
Cytokine Quantification by ELISA
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DuoSet ELISA to measure human sCD137 was from R&D Systems (Wiesbaden, Nordenstadt, Germany) and was performed as recommended by the distributor. Serum was diluted 1:2-fold for analysis. The concentration of IFN-γ in the cell culture supernatants was determined with the Human IFN-γ DuoSet ELISA kit (R&D Systems), according to the manufacturer's instructions.
The concentrations of TNF, IL-10, IL-13, and IL-17a in the cell culture supernatants were determined with human ELISA kits (Thermo Fisher Scientific, Waltham, MA), according to the manufacturer's instructions.
The concentrations of TNF, IL-10, IL-13, and IL-17a in the cell culture supernatants were determined with human ELISA kits (Thermo Fisher Scientific, Waltham, MA), according to the manufacturer's instructions.
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