4% formaldehyde-fixed SK-N-BE cells stained for Ankyrin brain (green) using ab131419 at 1/100 dilution in ICC/IF.

Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1/100 for 60 min at room temperature. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1/1000; 1/5000 for 60 min room temperature, 5 min room temperature.

Overlay histogram showing SH-SY5Y cells stained with ab131419 (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 (ab131419, 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 IgG1 [ICIGG1] (ab91353, 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 SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Western blot detection of Ankyrin brain in Rat brain membrane tissues using ab131419 at 1:1000 dilution.