Immunofluorescence analysis of Cytokeratin Pan (Cytokeratin 10, Cytokeratin 13, Cytokeratin 18, Cytokeratin 4, Cytokeratin 5, Cytokeratin 6 and Cytokeratin 8) was performed using 70% confluent log phase A-431 cells. The cells were fixed and permeabilized with ice-cold acetone at 4°C for 5 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin Pan Monoclonal Antibody (C-11) (Product # MA5-12231, 5 µg/mL) in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoskeletal localization. Panel e represents Raji cells showing no expression of cytokeratin. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.