Levels of IL-2 and IL-15 complex (IL-15 complexed with IL-15Rα) were analyzed using commercially available IL-2 ELISA (Invitrogen; 88-7025-22) and IL-15 complex ELISA (R&D Systems; DY6924), according to the manufacturer’s protocol. Cytokine plasma concentrations were quantified by electrochemiluminescence immunoassay using the U-Plex human proinflammatory Combo 1 panel MSD (Meso Scale Discovery) plates according to each manufacturer’s instructions. For ELISPOT immunoprecipitation, sterile clear plates (Millipore; MAIPS4510) were coated with anti-IFNγ monoclonal antibody (Thermo Fisher Scientific; M700A, 1:1,000 in PBS). PBMCs were plated at 100 µl/well (200,000 cells/well). Peptides were diluted in complete medium for final concentration of 2 µg/ml per well (JPT Innovative Peptide Solutions; PepMix HCMVA PM-PP65-1, and PepMix EBV PM-EBV-BARF1). In some wells, IL-15/IL-15Rα complex (IL-15 from Peprotech; 200-15, complexed with IL-15Rα from R&D Systems; 147-IR, 300 pg/ml) and IL-2 (Peprotech; 200-02, 100 pg/ml) were added and biotinylated anti-human IFNγ polyclonal antibody (Thermo Fisher Scientific; M701B, 1:1,000 in PBS/4% BSA) was added. Plates were scanned using an ELISPOT reader (Cellular Technology Limited; S6 Universal Analyzer) and analyzed using Cellular Technology Limited 5.2.12 Professional Analyzer software.
S6 universal analyzer
Manufactured by Cellular Technology
The S6 Universal Analyzer is a versatile laboratory instrument designed for the analysis of a wide range of samples. It features advanced technology to provide accurate and reliable results across various applications.
Automatically generated - may contain errors
Lab products found in correlation
Maips4510, by Merck Group (1 mentions)
Trueblue peroxidase substrate, by LGC (1 mentions)
Horseradish peroxidase conjugated donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Immunospot software, by Cellular Technology (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
Tissuelyser 2, by Qiagen (1 mentions)
2 protocols using s6 universal analyzer
1
Quantifying Cytokine Levels in PBMCs
2
CCHFV Antigen Detection Assay
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Mouse tissues were dissected into tubes containing a steel bead and SW13 growth media and homogenized at 30hz using a TissueLyser II (Qiagen). Clarified tissue homogenates were diluted in L-15 media and then applied to SW13 cells in a 96-well format. Sample was then removed, and cells overlaid with L-15 media supplemented with 0.8% methylcellulose (Sigma). Cells were incubated at 37 °C for 40 h. Overlay was removed, and cells fixed with 2% paraformaldehyde overnight at 4 °C. To detect CCHFV antigen, mouse anti-CCHFV Gc monoclonal 11E7 (BEI Resources) was applied at 500 ng/mL. Plates were washed, and bound antibody detected with horseradish peroxidase conjugated donkey anti-mouse IgG (Jackson Immuno Research). Plates were developed with TrueBlue peroxidase substrate (Seracare) and foci counted with an S6 Universal analyzer and Immunospot software (Cellular Technology Limited).
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