Levels of IFN-γ in supernatants from T cell clone recognition assays were measured by ELISA using the ELISA MAX Deluxe Set Human IFN-g kit (Biolegend) following the manufacturer’s instructions. Concentrations were determined by interpolating from a standard curve.
Elisa max deluxe set human ifn γ kit
Manufactured by BioLegend
Sourced in United States
The ELISA MAX™ Deluxe Set human IFN-γ kit is a laboratory tool used to detect and quantify human interferon-gamma (IFN-γ) levels in various sample types. It is a sensitive and reliable enzyme-linked immunosorbent assay (ELISA) kit that provides the necessary components to perform the analysis.
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8 protocols using elisa max deluxe set human ifn γ kit
1
Quantifying IFN-γ in T-cell Assays
2
Quantifying IFN-γ in T-cell Assays
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Levels of IFN-γ in supernatants from T cell clone recognition assays were measured by ELISA using the ELISA MAX Deluxe Set Human IFN-g kit (Biolegend) following the manufacturer’s instructions. Concentrations were determined by interpolating from a standard curve.
3
Quantifying IFNγ Production in P-BCMA-ALLO1 Assay
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To measure IFNγ production from P-BCMA-ALLO1, the ELISA MAX Deluxe Set Human IFNγ kit (catalog #430104, BioLegend) was run according to manufacturer’s instructions. Supernatants from the cytotoxicity assay were thawed on ice and mixed before diluting 100-fold and applying to coated ELISA plates. Developed ELISA plates were read on the Biotek Synergy in absorbance mode at 450 and 570 nm. The absorbance at 450 nm in each well is directly proportional to the amount of IFNγ in the supernatant.
4
Measuring Human IFN-γ by ELISA
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Measurement of human IFN-γ in the culture supernatant was performed with ELISA MAX™ Deluxe Set human IFN-γ kit (BioLegend) according to manufacturer’s instruction. Cell-free supernatant was harvested from cell culture 24 h after coculture with effectors (E:T ratio 1:2). The plate was then washed, incubated with tetramethylbenzidine substrate (Agilent Technologies, CA), and read at 450 nm using a Synergy HT microplate reader (Biotek, VT).
5
Measuring Human IFN-γ by ELISA
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Measurement of human IFN-γ in the culture supernatant was performed with ELISA MAX™ Deluxe Set human IFN-γ kit (BioLegend) according to manufacturer’s instruction. Cell-free supernatant was harvested from cell culture 24 h after coculture with effectors (E:T ratio 1:2). The plate was then washed, incubated with tetramethylbenzidine substrate (Agilent Technologies, CA), and read at 450 nm using a Synergy HT microplate reader (Biotek, VT).
6
Antibody Inhibition of IFN-γ Release
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The ability of antibody hits to inhibit IFN-γ release from activated T cells in human PBMCs was assayed using the ELISA MAX ™ Deluxe Set Human IFN-γ Kit (BioLegend, #430104). Human PBMCs, isolated T cells or isolated NK cells were seeded at 200,000 cells per well in 96-well plates and incubated for 30 min at 37°C with anti-A 2A R antibodies, ZM-241385, or AB928 (MedChemExpress, # HY-129393), after which 1, 3, or 10 μM NECA was added for another 30 min. Dynabeads ® Human T-Activator CD3/CD28 (Invitrogen #11161D) were then added at 1 μl of beads per 4 x 10 6 cells and incubated for 3 d at 37°C to activate T cells. Finally, culture media was harvested and used directly for the IFN-γ ELISA according to the manufacturer's instructions, and cells were stained with activation markers to determine activation level by flow cytometry.
7
Quantifying IFN-γ in Cell Cultures
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Interferon-γ (IFNγ) in culture supernatants was quantified with the ELISA MAX™ Deluxe Set Human IFN-γ Kit (BioLegend; San Diego, CA, USA) according to the manufacturer’s recommendations.
8
Measuring Plasma Cytokine Levels
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Plasma cytokine levels were determined using the ELISA MAX Deluxe Set Human IFN-γ Kit (BioLegend) according to the manufacturer’s recommendations. Plasma was diluted 1:1 or 1:10 in 1× Assay Diluent A. Optical density was read at 450 nm and 570 nm using a SpectraMax i3x platform (Molecular Devices).
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