Overlay histogram showing MCF7 cells stained with ab221256 (red line). The cells were fixed with 80% methanol (5 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 (ab221256, 1/50 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) PerCP (ab222107) 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 50 mW Blue laser (488nm) and 685/35 bandpass filter.

This antibody gave a positive signal in MCF7 cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.