Splenocytes from mice at up to 5 × 106 cells were seeded together with 5 mg/mL mouse IL-2 (Biolegend), 1 μL BD GolgiPlug (Becton Dickinson, Franklin Lakes, NJ) or Brefeldin A (ThermoFisher Scientific) and 10 μg peptide in 96-well tissue culture plates, followed by incubation at 37°C, 5% CO2 for 5 h. For Plasmodium studies, the mouse IL-2 addition was omitted. They were then stained with Zombie Aqua Fixable Viability kit (Biolegend) for 30 min, washed and followed by antibody stainings for 30 min, selected from the following panel: purified CD16/32 (clone 93); CD3 PE/Dazzle 594 (clone 17A2); CD3ε Brilliant Violet 421 (clone 145-2C11); CD4 APC/Cy7 (clone GK1.5); CD8a PerCP/Cy5.5 (clone 53-6.7); CD8b PerCP/Cy5.5 (clone YTS 156.7.7, all from Biolegend). After washing, cells are fixed in 4% formaldehyde, permeabilized with 0.1% saponin (Sigma-Aldrich) or per buffer (BD Bioscience), followed by staining with IFNγ FITC (clone XMG1.2, Biolegend/eBioscience).
Purified cd16 32 clone 93
Manufactured by BioLegend
Purified CD16/32 (clone 93) is a mouse monoclonal antibody that binds to the Fc gamma receptor III and II (CD16 and CD32). It is a useful tool for blocking Fc gamma receptor-mediated functions in mouse cells.
Automatically generated - may contain errors
Lab products found in correlation
Golgiplug, by BD (1 mentions)
Saponin, by Merck Group (1 mentions)
Zombie aqua fixable viability kit, by BioLegend (1 mentions)
Brefeldin a, by Thermo Fisher Scientific (1 mentions)
Mouse il 2, by Thermo Fisher Scientific (1 mentions)
Cd3 pe dazzle 594 clone 17a2, by BioLegend (1 mentions)
Cd4 apc cy7 clone gk1, by BioLegend (1 mentions)
Mouse il 2, by BioLegend (1 mentions)
Ifnγ fitc clone xmg1, by BioLegend (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Dspe peg2000, by Avanti Polar Lipids (1 mentions)
Collagenase dispase, by Roche (1 mentions)
Dotap, by Avanti Polar Lipids (1 mentions)
Dnase 1, by Roche (1 mentions)
Brefeldin a, by BioLegend (1 mentions)
Interleukin 2 (il 2), by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Mouse il 1β, by R&D Systems (1 mentions)
Tnf α duoset elisa kits, by R&D Systems (1 mentions)
2 protocols using purified cd16 32 clone 93
1
Murine Splenocyte Cytokine Profiling
2
Lipid-based Nanoparticle Formulations for Immunological Studies
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DOTAP, 1,2-distearoryl-snglycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol-2000) ammonium salt (DSPE-PEG2000) were purchased from Avanti Polar Lipids (Alabaster, AL). Cholesterol and protamine sulfate salt from salmon (Grade X) were purchased from Sigma-Aldrich (St. Louis, MO). Collagenase/dispase and DNase I were purchased from Roche diagnostics. Fluorescent antibodies against mouse B220 (clone RA3-6B2), CCR7 (clone 4B12), CD4 (clone GK1.5), CD8a (clone 53–6.7), CD11b (clone M1/70), CD11c (clone N418), CD44 (clone IM7), CD62L (clone MEL-14), CD69 (clone H1.2F3), CD80 (clone 16-10A1), CD86 (clone GL-1), CD103 (clone 2E7), CD127 (clone A7R34), CD138 (clone 281–2), GL-7 (clone GL7), IFN-γ (clone XMG1.2), IL-2 (clone JES6-5H4), Ki67 (clone SolA15), KLRG1 (clone 2F1/KLRG1), Ly6G (clone 1A8), MHC-II (clone M5/114.15.2), NK1.1 (clone PK136), PD-1 (clone 29F.1A12), PDCA1 (clone 927), TIM-3 (clone 8B.2C12), TNF-α (clone MP6-XT22), purified CD16/32 (clone 93), and brefeldin A were from BioLegend (San Diego, CA). PNA and Lipofectamine 2000 were from Thermo Fisher Scientific. The mouse IL-1β, IL-2, IL-6, IL-10, IFN-γ, and TNF-α DuoSet ELISA kits were purchased from R&D Systems (Minneapolis, MN).
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