Flow cytometry was performed on single cell suspensions of splenic, iliac lymph node, bladder and prostate tissues. Tissues were made into single cell suspensions by passing through 40 μm mesh filter membranes to remove debris and washing with 2% FCS (HyClone) in PBS (Gibco). Samples were then fixed and permeabilized using fixation-permeabilization buffers (eBioscience Cat. Numbers 8222-49 and 8333-56), according to manufacturer’s instructions. Following this cells were stained with the following mouse antibodies (Ly6G-FITC, Ly6C-PE, IL4-PE, B220-PE, CD3-PerCp, CD86-PerCp, CD11b-APC, IL17A-APC (eBioscience), CD8-FITC, IFNγ-FITC, CD4-PerCp, CD4-APC (Biolegend)), and run on an Accuri benchtop C6 cytometer. Results were analyzed using FlowJo™ software and statistics generated using Prism™ software from GraphPad.
Cd86 percp
Manufactured by Thermo Fisher Scientific
Sourced in United States
CD86-PerCp is a fluorescent-conjugated antibody that binds to the CD86 surface antigen. CD86 is a costimulatory molecule expressed on antigen-presenting cells that plays a role in T cell activation. The PerCp (peridinin-chlorophyll protein) fluorescent label allows for detection and quantification of CD86-expressing cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cd3 percp, by Thermo Fisher Scientific (1 mentions)
B220 pe, by Thermo Fisher Scientific (1 mentions)
Cd11b apc, by Thermo Fisher Scientific (1 mentions)
Cd8 fitc, by BioLegend (1 mentions)
Cd4 apc, by BioLegend (1 mentions)
Ly6c pe, by Thermo Fisher Scientific (1 mentions)
Cd4 percp, by BioLegend (1 mentions)
Ly6g fitc, by Thermo Fisher Scientific (1 mentions)
Il 17a apc, by Thermo Fisher Scientific (1 mentions)
Il 4 pe, by Thermo Fisher Scientific (1 mentions)
Fixation permeabilization buffer, by Thermo Fisher Scientific (1 mentions)
Ifn γ fitc, by BioLegend (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Prism software, by GraphPad (1 mentions)
Ow cytometry, by BD (1 mentions)
Tryple, by Thermo Fisher Scientific (1 mentions)
Cd163 apc, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
2 protocols using cd86 percp
1
Flow Cytometric Analysis of Immune Cells
2
Macrophage Polarization Assay with sEVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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RAW264.7 macrophages were stimulated with 500 ng/mL LPS (Sigma‒Aldrich, MO, USA) in the presence and absence of 30 μg/mL Nor-sEV and Hypo-sEV for 12 h. Then, RAW264.7 cells were washed with PBS, treated with TrypLE (Gibco, USA) and resuspended in 0.5 mL of PBS supplemented with 5% FBS. Macrophage surface marker expression was analyzed by flow cytometry (BD Bioscience, San Jose, CA, USA) after the samples were stained with CD86-PerCP, CD163-APC and CD206-FITC monoclonal antibodies (eBioscience, San Diego, CA, USA) for 30 min in the dark at 4 °C.
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