Overlay histogram showing Hela cells stained with ab134093 (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 (ab134093, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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 HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Anti-MUC16 antibody [EPSISR23] (ab134093) at 1/200000 dilution + NIH: OVCAR-3 (Human ovary adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Secondary
Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 1519 kDa

All lanes : Anti-MUC16 antibody [EPSISR23] (ab134093) at 1/10000 dilution

Lane 1 : Human ovary cancer lysate
Lane 2 : Human uteruslysate
Lane 3 : NIH:OVCAR-3 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 1519 kDa

Immunofluorescent analysis of OVCAR-3 cells labelling MUC16 with ab134093 at 1/100 dilution.