Overlay histogram showing HeLa cells (negative cell line (left)) and HL-60 cells (positive cell line (right)) stained with ab225476 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal Goat serum to block non-specific protein-protein interactions followed by the antibody (ab225476) (1x 10⁶ cells in 100 µl at 0.08 µg/ml (1/6250 dilution)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Allophycocyanin (ab232814) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 670/14 bandpass filter.

This antibody gave a positive signal in HL-60 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.