Overlay histogram showing RAW264.7 cells treated (red line) or untreated (green line) with 100 ng/mL of lipopolysaccharides (LPS) for 7 hours with addition of 1 µg/mL brefeldin A (BFA) for the last 3 hours and subsequent staining with ab237355. The cells were fixed with 4% formaldehyde and then permeabilized with 90% methanol. The cells were then incubated in 1x PBS/10% normal serum to block non-specific protein-protein interactions followed by the antibody (ab237355) (1x10⁶ in 100µl at 1/50 dilution (10 µg/ml)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20 mW Blue laser (488 nm) and 585/42 bandpass filter.