Fitc conjugated anti human cd83

In vitro generated DCs were assessed for surface expression using fluorochrome-conjugated monoclonal antibodies: PE conjugated anti-human CD1a, PE conjugated anti-human CD11c, FITC conjugated anti-human CD83 and APC conjugated anti-human HLA-DR (BD Biosciences, San Jose, USA). DCs were harvested and washed with cold PBS, and antibodies were added to bind at 4°C for 20 minutes in the dark. Then cells were washed twice with PBS for flow cytometry, performed with FACSCanto (BD Biosciences) flow cytometer. Data were analyzed with FlowJo software.

For BMDCs and iPSDCs staining, the following monoclonal antibodies were used: PE-conjugated anti-human CD11c, FITC-conjugated anti-human CD80, FITC-conjugated anti-human CD83, PE-conjugated anti-human CD40, PE-conjugated anti-human HLA-ABC, PE-conjugated anti-human HLA-DR (all from BD Biosciences), primary antibodies against HLA-A were Anti-HLA A antibody, and secondary antibodies were Goat Anti-Rabbit IgG H&L (FITC) (all from Abcam). For CTOSs staining, the following monoclonal antibodies were used: FITC anti-human EpCAM (Miltenyi), PE anti-human CD45 (BD Biosciences). Primary antibodies against CEA were anti-human CEA monoclonal antibody clone COL-1 (Leica Biosystems), and secondary antibodies were FITC-conjugated anti-mouse IgG polyclonal antibody (Agilent Technologies).