Supernatants from cultured DCs were harvested and analyzed for secreted cytokines as indicated below unless otherwise noted. For IFNγ release, supernatants from T cells cultured in the presence or absence of treated DCs were analyzed using ELISA MAX kits [Biolegend (San Diego, CA)]. To measure nitrite production, supernatants were subjected to the Griess Test assay [Invitrogen (Waltham, MA)]. To study cytokine production, supernatants were analyzed using the LEGENDplex system [Biolegend (San Diego, CA)], followed by flow cytometric analysis using Biolegend software.
Legendplex system
Manufactured by BioLegend
Sourced in United Kingdom, Germany
The LEGENDplex system is a multiplex assay platform designed for the simultaneous detection and quantification of multiple analytes in a single sample. The system utilizes fluorescence-coded beads and flow cytometry technology to enable high-throughput and efficient analysis of various biomolecules, including cytokines, chemokines, and other proteins.
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Lab products found in correlation
4 protocols using legendplex system
1
Cytokine and Nitrite Analysis of Cultured DCs
2
Measuring CXCL9 and CXCL10 in Plasma
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Levels of CXCL9 and CXCL10 were measured in plasma (diluted twofold) using the mouse cytokine release syndrome panel LEGENDplex system (BioLegend, UK), according to the manufacturer’s instructions. The acquisition was performed on the BD LSRFortessa.
3
Multiplex Cytokine and Chemokine Profiling
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Cytokine and chemokine profile was characterized using the LEGENDplex™ system (BioLegend). A more detailed protocol is published (30 (link)). Briefly, we used the Mouse Inflammation Panel (13-plex) system. Serum from WT and TKO mice was collected and incubated with fluorescence-encoded capture beads to cytokine and chemokine targets including CCL2, TNF and IFNγ. The fluorescent signals of analyte-specific bead regions were quantified using flow cytometry, and the concentrations of particular analytes were determined using provided data analysis software (BioLegend, LegendPlex™ software v8.0).
4
Characterization of Cytokine and Chemokine Profile
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Cytokine and chemokine profile was characterized using the LEGENDplex™ system (BioLegend, Koblenz, Germany). A more detailed protocol is published (35 (link)). Briefly, we used the Mouse Inflammation Panel (13-plex). Serum isolated from WT infected mice was incubated with fluorescence-encoded capture beads to cytokine and chemokine targets, including TNF, IL-1β, IL-6, and IFNγ. The fluorescent signal of analyte-specific bead regions was quantified using flow cytometry, and the concentrations of particular analytes were determined using provided data analysis software (BioLegend, LegendPlex™ software v8.0).
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