Flow cytometry overlay histogram histogram showing wild-type (green line) and CD1C knockout Jurkat cells stained with ab273049 (red line). The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab273049) (1x10⁶ in 100 µl at 0.2 µg/ml) for 30 min at 4°C.

The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.

Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type Jurkat - black line; CD1C knockout Jurkat - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD1c with ab273049 at 1/500 dilution (0.1ug) (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Cells were stained with rabbit IgG (Left) or ab273049 (Right). Then stained with anti-CD19 conjugated to Alexa Fluor^® 647.

Gated on viable cells.

Flow cytometric analysis of THP-1 (Human monocytic leukemia monocyte)(Left) / Jurkat (Human T cell leukemia T lymphocyte)(Right) cells labelling CD1c with ab273049 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control: THP-1 (PMID: 24935257).

Gated on viable cells.