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Duoset human ifnγ elisa kit

Manufactured by R&D Systems
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The DuoSet Human IFNγ ELISA kit is a sandwich enzyme-linked immunosorbent assay (ELISA) designed for the quantitative measurement of human interferon gamma (IFNγ) in cell culture supernatants, serum, and plasma samples.

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Glo substrate, by R&D Systems (1 mentions)

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DuoSet ELISA kits (1131 citations) DuoSet ELISA (635 citations) ELISAs (144 citations) DuoSets (49 citations) IFNγ ELISA (12 citations) IFNγ Duoset Elisa (2 citations) Human ELISA DuoSet (2 citations) Human kits (2 citations)

3 protocols using duoset human ifnγ elisa kit

1

IFNγ Cell-based ELISA Protocol

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Cell-based ELISAs were carried out as previously published.44 Target cells, effector T cells (104/well) and/or peptide, or IFNγ standards, were plated in 384-well plates after overnight plate coating with IFNγ capture antibody. After 48 h, the plates were washed and developed for IFNγ cell-ELISA, per manufacturer’s instructions (R&D DuoSet Human IFNγ ELISA kit), with the use of a luminescent HRP substrate (Glo Substrate, R&D Systems). Luminescence was measured using a FLUOstar Omega plate reader.
Sanderson J.P., Crowley D.J., Wiedermann G.E., Quinn L.L., Crossland K.L., Tunbridge H.M., Cornforth T.V., Barnes C.S., Ahmed T., Howe K., Saini M., Abbott R.J., Anderson V.E., Tavano B., Maroto M, & Gerry A.B. (2019). Preclinical evaluation of an affinity-enhanced MAGE-A4-specific T-cell receptor for adoptive T-cell therapy. Oncoimmunology, 9(1), 1682381.
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2

Quantifying IFN-γ Production in CIK Cells

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To determine the amount of IFN-γ produced by CIK cells, a Duoset® Human IFN-γ ELISA kit (R&D Systems) was used, as described by the manufacturer. The final measurements were performed at 450 and 540 nm wavelengths and the data were analyzed using MARS data analysis (BMG Labtech).
Li Y., Sharma A., Bloemendal M.W., Schmidt-Wolf R., Kornek M, & Schmidt-Wolf I.G. (2021). PD-1 blockade enhances cytokine-induced killer cell-mediated cytotoxicity in B-cell non-Hodgkin lymphoma cell lines. Oncology Letters, 22(2), 613.
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3

NKTs and CD8+ T Cell-Mediated Tumor Killing

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NKTs and CD8+ T cells (105 cells/well) were cocultured with tumor cell lines (M14-wt, M14-A2, C8161 and SK-MEL-5) (105 cells/well) at an E:T ratio of 1:1 in 24-well plates, in complete medium, in the absence of cytokines. After 3 days of culture, cells were harvested and stained for CD3 (APC-H7, clone SK7 from BD Biosciences) and CD276 (BV421, clone 7–517 from BD Biosciences) monoclonal (m)Abs to detect T cells and tumor cells, respectively. Percentage of residual tumor cells in culture were enumerated by flow cytometry. NKTs (105 cells/well) were also cocultured with HLA-A2+ T cell blasts loaded with the YMDGTMSQV peptide (100 nM) at an E:T ratio of 1:1 or activated with 10 µL of anti-Vα24 (clone 6B11, from BD Biosciences) in 1 mL of complete medium in the absence of cytokines. Culture supernatants were harvested after 24 hours of culture and IFNγ measured in 100µl of supernatant with the DuoSet Human IFNγ ELISA kit (R&D System). Data acquisition was performed on a Synegrgy2 microplate reader (BioTek) using the Gen5 software.
Landoni E., Smith C.C., Fucá G., Chen Y., Sun C., Vincent B.G., Metelitsa L.S., Dotti G, & Savoldo B. (2019). A High Avidity T-Cell Receptor Redirects Natural Killer T-Cell Specificity and Outcompetes the Endogenous Invariant T-Cell Receptor. Cancer immunology research, 8(1), 57-69.
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