NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling ATP7A using ab131400 at 1/100 dilution in ICC/IF. Cells were fixed using 4% formaldehyde for 15 minutes at room temperature. Incubation with primary antibody was performed for 1 hour at room temperature. Secondary antibody used was a goat anti-mouse ATTO 488 (green) at 1/200 dilution for 1 hour at room temperature. Counterstained with Phalloidin Texas Red F-actin stain. Nuclei were stained with DAPI (blue).

Overlay histogram showing HT1080 cells stained with ab131400 (red line). The cells were fixed with 4% paraformaldehyde (10 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 followed by the antibody (ab131400, 0.1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HT1080 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.