α tubulin primary antibody

Cells grown in two-well chamber slides were fixed with 10% neutral buffered formalin, permeabilized with 0.2% Triton X-100 in PBS, and blocked with 5% goat serum, 1% bovine serum albumin (BSA), and 0.5% Tween-20 in PBS. The cytoplasm was stained by an overnight incubation with α-tubulin primary antibody (Cell Signaling Technology #2125) at 4 °C, and then a 2 h, room temperature incubation with goat anti-rabbit IgG Alexa Fluor 488 antibody. Coverslips were mounted using ProLong Gold Antifade Mountant with DAPI to stain nuclei. Slides were imaged on a Zeiss Axio Imager 2 with a 63× objective. Mitotic catastrophe was scored in 50–100 cells per treatment group. Cells were scored positive for mitotic catastrophe if they contained micronuclei, one nucleus with two or more lobes, or multiple nuclei^{17 (link)}. Data presented are the mean ± SEM for three independent experiments. Statistical significance was calculated by Student’s t-test.

Cal33 cells were seeded and grown on glass coverslips and exposed to DMSO-only, 100 nM, 500 nM or 1 μM albendazole treatments for 24 hours. Three replicates of each treatment were prepared. Cells were fixed to the coverslips using 100% methanol (pre-chilled at −20°C). Samples were blocked using 5% normal donkey serum (Wisent) in PBS. Cells were then incubated for 1 hour with α-tubulin primary antibody (1:50, Cell Signaling Technology Cat. ID 2125) at room temperature, then washed extensively with PBS. Alexa Fluor^® 488 goat anti-rabbit IgG secondary antibody (1:1000, Molecular Probes, Inc.A-11008) was added to each slide and then incubated in the dark for 60 minutes. DAPI (Molecular Probes, Inc., 30 nM) was used for nuclear staining. Images were obtained under 40x magnification using an Olympus Provis AX70 microscope.