Western blot detection of Shank1 in Rat brain membrane lysates using ab94576 at 1:1000 dilution.

paraffin-embedded, bouin's-fixed mouse backskin stained for SHANK1 using ab94576 at 1/100 dilution in immunohistochemical analysis. FITC Goat Anti-Mouse (green) at 1/50 dilution was used as the secondary antibody. Filaggrinlike staining (upper layer aggregations of staining) is observed.

Overlay histogram showing SH-SH5Y cells stained with ab94576 (red line). The cells were fixed with 80% methanol (5 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 (ab94576, 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 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.