Flow cytometric analysis of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with with a Myc-tagged human TNFRSF14/HVEM expression construct labeling TNFRSF14/HVEM with ab224708 at 1/400 dilution (red) compared with a rabbit monoclonal IgG Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

Flow cytometric analysis of human PBMCs (peripheral blood mononuclear cells) labeling TNFRSF14/HVEM with ab224708 at 1/400 dilution (red) compared with a rabbit monoclonal IgG Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081), at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with with a Myc-tagged human TNFRSF14/HVEM expression construct labeling TNFRSF14/HVEM with ab224708 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in 293T cells transfected with a Myc-tagged human TNFRSF14/HVEM expression construct. The nuclear counter stain is DAPI (blue). Cells are counter stained for Myc using an Anti-Myc tag antibody [9E10] (Alexa Fluor^® 594) (ab223894) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.