Immunofluorescence Staining of Tubulin in Monolayer Cells

Monolayer culture was performed by seeding cell on a glass bottom culture dish with 60% confluence. After that, cells were prepared for immunofluorescence staining according to standard protocols. Cells were fixed in cold 4% paraformaldehyde for 20 min, washed 3 times in 0.5% Triton X-100 in PBS (PBST) for 5 min, and incubated in blocking solution consisting of 1% bovine serum albumin (BSA) in PBST for 2 h. Anti-alpha tubulin (Abbkine, diluted 1:200) and DyLight549 goat anti-mouse (Abbkine, diluted 1:200) antibodies were used as the primary antibody and secondary antibody, respectively. 4′,6-diamidino-2-phenylindole (DAPI) was used for nuclear staining. A laser confocal microscope (NIKON Elipse Ti) was used for observation [16 (link), 17 (link)].