Overlay histogram showing HepG2 cells treated with TSA (500ng/ml, 4hr) stained with ab214511 (red line) and untreated HepG2 cells stained with ab214511 (green line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab214511, 1/50 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (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 blue solid-state laser (488nm) and 585/42 bandpass filter.

ab214511 staining p53 (acetyl K373) in HepG2 cells +/- treatment with Etoposide (25μM, 5 hours) and Trichostatin A (500ng/ml, 4 hours). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 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 ab214511 at 1/50 dilution (pseudocolored in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

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