Immunofluorescence analysis of beta tubulin was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with beta Tubulin Loading Control Monoclonal Antibody (BT7R), Alexa Fluor 555 (Product # MA5-16308-A555) at 1:25 dilution in 0.1% BSA and incubated at 4 degree Celsius overnight (Panel a: red). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: green) was stained with Alexa Fluor® 488 Phalloidin (Product # A12379, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the isotype control. The images were captured at 60X magnification.