CD4+ T cells and CD8+ T cells from the prepared TILs were stained with FITC-conjugated anti-CD3 Ab (BD Pharmingen), PE-conjugated anti-CD4 Ab, and PE-conjugated anti-CD8 Ab (BD Pharmingen). DCs from the prepared TILs were stained with FITC-conjugated anti-CD11c Ab (BD Pharmingen), PE-conjugated anti-CD80Ab (BD Pharmingen), and PE-Cy5-conjugated anti-CD86Ab (BioLegend) as described previously [48 (link)]. Macrophages from the prepared TILs were stained with FITC-conjugated anti-F4/80 Ab (BD Pharmingen), PE-conjugated anti-CD80Ab (BD Pharmingen), and PE-Cy5-conjugated anti-CD86Ab (BioLegend). The cells were analyzed by flow cytometry, as described above.
Pe cy5 conjugated anti cd86 ab
Manufactured by BioLegend
The PE-Cy5-conjugated anti-CD86 Ab is a monoclonal antibody that binds to the CD86 cell surface protein. The antibody is conjugated to the PE-Cy5 fluorescent dye, which can be detected using flow cytometry.
Automatically generated - may contain errors
3 protocols using pe cy5 conjugated anti cd86 ab
1
Multicolor Flow Cytometry Analysis of TILs
2
Characterization of Tumor-Draining LNs
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Tumor-draining LNs were harvested from mice treated with E7-specific protein vaccine and/or anti-PD-L1 Ab 7 days after the last protein vaccination (on day 28 after tumor challenge) (Figure 4 A). Single cell suspensions were prepared as described previously [47 (link)]. To detect the maturation status of DCs and the M1-like polarization of macrophages, the cells were stained with FITC-conjugated anti-CD11c Ab, FITC-conjugated anti-F4/80 Ab, PE-conjugated anti-CD80 Ab, and PE-Cy5-conjugated anti-CD86 Ab (BioLegend) or PE-conjugated anti-MHC class I and PE-Cy5-conjugated anti-MHC class II Abs (BioLegend). The cells were analyzed by flow cytometry, as described above.
3
Evaluating anti-PD-L1 Ab's effect on DC maturation
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To determine whether anti-PD-L1 Ab enhanced DC maturation, BMM-derived DCs treated with 50 ng/mL LPS were used as a positive control. DCs were treated with or without LPS and anti-PD-L1 Ab (50 μg/mL) in a 5% CO2 atmosphere at 37 °C overnight. The cells were stained with FITC-conjugated anti-CD11c Ab (BioLegend), PE-conjugated anti-CD80 Ab (BioLegend), anti-MHC class I Ab (Biolegend), PE-Cy5-conjugated anti-CD86 Ab (BioLegend) or anti-MHC class II Ab (Biolegend). The cells were analyzed by flow cytometry, as described above.
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