Overlay histogram showing Hela cells stained with DNM2 (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 (DNM2, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight 488 Conjugated Highly Cross-Adsorbed(OH191631) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG (1ug/1x10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.

All lanes: Anti-DNM2 Antibody (N-terminal) at 1:2000 dilution Lane 1: Hela whole cell lysate Lane 2: K562 whole cell lysate Lane 3: NIH/3T3 whole cell lysate Lane 4: mouse brain lysate Lane 5: Jurkat whole cell lysate Lysates/proteins at 20ug per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10,000 dilution. Predicted band size: 98kD Blocking/Dilution buffer: 5% NFDM/TBST.