Anti cd11c fitc apc

On day 10, LPS-stimulated cDCs and unstimulated cells were harvested by centrifugation and resuspended in PBS. An average of 5 x 10 5 cells per stain was subjected to subsequent analyses. Prior to staining, Fc receptors on cDCs were blocked by preincubation with anti-CD16/ CD32 antibodies (Biolegend) for 5-10 min on ice. Surface staining was performed by incubating the cells with fluorochrome-conjugated specific antibodies, or the corresponding isotype controls, for at least 20 min in dark on ice. The following antibodies and isotype controls (unless stated otherwise, all purchased from Miltenyi Biotec, Bergisch Gladbach, Germany) were employed: anti-CD11c-FITC/-APC (#130-102-466/-493), anti-MHC-II-APC (#130-102-139), anti-CD40-PE (#130-102-599), anti-CD80-PE (#130-102-613), anti-CD83-PE (#130-104-474), anti-CD86-APC (#130-102-558), anti-hamster IgG-FITC/-PE/-APC (eBiosciences, San Diego, CA, USA), anti-rat IgG2a-PE (eBiosciences), anti-rat IgG2b-APC (Immunotools), and REA Control. Flow cytometric analyses were performed on a FACSCalibur (BD Biosciences). A total of 20,000 events per sample were acquired, and data were evaluated using the CellQuestPro software (BD Biosciences).