Flow cytometric analysis of permeabilized Jurkat cells using ab109255 (red) or a rabbit IgG (negative) (green).

Overlay histogram showing HeLa cells stained with ab109255 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab109255, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was goat anti-rabbit DyLight® 488 (IgG H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Flow cytometry analysis of Plasmacytoid dendritic cells, staining IRF7 with ab109255. Cells were fixed in paraformaldehyde and fixed in saponin. The sample was incubated with the primary antibody (1/100 in 2% human serum + 0.5 mM EDTA) for 20 minutes at 4°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/100) was used as the secondary antibody.Gating Strategy: FSC/SSC lymphocytes and then FSC-A/FSC-H singletsSee Abreview

ab109255 was used to detect IRF7 in Human tonsil tissue using Immunohistochemistry (Frozen sections). Human tonsil tissue was frozen and fixed with acetone fixed, sample permeabilized with 0.05% Tween and blocked with 6% BSA for 1 hr at room temp. Primary antibody was diluted 1/100 in 2% BSA in TBS/Tween and incubated for 12 hr at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal(1/25) was used as the secondary antibody. IRF7 staining in green, DAPI in blue. See Abreview