Anti beta Tubulin  

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 (mouse myoblast cell line) cells labeling beta Tubulin with M32657 at 1/25 dilution, followed by DylightR 488-conjugated goat anti-rabbit IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on C2C12 cell line. The nuclear counter stain is DAPI (blue).

All lanes : Anti-beta Tubulin at 1:2000 dilution
Lane 1: C2C12 whole cell lysate
Lane 2: L6 whole cell lysate
Lane 3: mouse spleen lysate
Lane 4: mouse thymus lysate
Lane 5: NIH/3T3 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.

Overlay histogram showing NIH/3T3 cells stained with M32657 (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 (M32657, 1:25 dilution) for 60 min at 37oC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLightR 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at 37oC. Isotype control antibody (blue line) was rabbit IgG (1g/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.