Overlay histogram showing Hela cells stained with AP2512D (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP2512D, 1:25 dilution) for 60 min at 37?C. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight? 488 Conjugated Highly Cross-Adsorbed(NA168821) at 1/400 dilution for 40 min at 37?C. Isotype control antibody (blue line) was rabbit IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Overlay histogram showing Hela cells stained with AP2512D (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP2512D, 1:25 dilution) for 60 min at 37?C. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight? 488 Conjugated Highly Cross-Adsorbed(NA168821) at 1/400 dilution for 40 min at 37?C. Isotype control antibody (blue line) was rabbit IgG (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

All lanes : Anti-EZH2 Antibody at 1:2000 dilution Lane 1: Hela whole cell lysate Lane 2: Jurkat whole cell lysate Lane 3: MCF-7 whole cell lysate Lane 4: Neuro-2a whole cell lysate Lane 5: T47D whole cell lysate Lysates/proteins at 20 μg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 85 kDa Blocking/Dilution buffer: 5% NFDM/TBST.