Overlay histogram showing HEK293 cells stained with ab91205 (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 (ab91205, 1/100 dilution) 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 HEK293 cells fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

ab91205, at a 1/200 dilution, staining eNOS in paraffin embedded Human lymph node (left) and colon cancer (right) tissues by Immunohistochemistry. Detection was by DAB staining.

ab91205, at a 1/200 dilution, staining eNOS in paraffin embedded Human stomach cancer (left) and ovary cancer (right) tissues by Immunohistochemistry. Detection was by DAB staining.