Overlay histogram showing C2C12 cells stained with BD-PB3741(green line). The cells were fixed with 2% paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then incubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at Room temperature. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (mouse myoblast cell line) cells labeling beta II Tubulin with BD-PB3741 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-rabbit IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoskeleton staining on C2C12 cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin at 1/100 dilution (red). The nuclear counter stain is DAPI (blue).

All lanes : Anti-beta II Tubulin Antibody at 1:2000 dilution Lane 1: C2C12 whole cell lysate Lane 2: NIH/3T3 whole cell lysate Lane 3: Mouse brain lysate Lane 4: Human brain lysate Lane 5: Hela whole cell lysate Lane 6: A431 whole cell lysate Lane 7: 293 whole cell lysate Lane 8: MCF-7 whole cell lysate Lane 9: PC-12 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 : 50 kDa Blocking/Dilution buffer: 5% NFDM/TBST.