Recombinant PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab209478)

Immunofluorescent analysis of HeLa (human cervical cancer) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes, blocked with Image-iT^® FX Signal Enhancer at room temperature for 30 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209478 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/50 dilution (showing no signal) and ab195884, Rat monoclonal [YOL1/34] to Tubulin Microtubule Marker (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Flow cytometry overlay histogram showing HeLa cells stained with ab209446 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% Methanol for 30 min at -20°C. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab209446) (1x10⁶ in 100µL at 0.01µg/mL, 0.1µg/mL, 1µg/mL (1/50000, 1/5000, 1/500)) for 30 min at 22°C.

Isotype control antibody (black line) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

Overlay histogram showing HeLa cells stained with ab208741 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab208741, 1/500 dilution) for 30 min at 22ºC.

Isotype control antibody (black line) was rabbit IgG monoclonal [EPR25A] (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20 mW Solid State Blue Laser (488nm) and 585/40 bandpass filter.