Immunofluorescence analysis of Cytokeratin 17 was performed using MCF 10A and MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin 17 Monoclonal Antibody (Product # MA1-06325) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32766) at a dilution of 1:2000 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). Panel d represents the composite image showing cytoskeletal localization of Keratin 17 in MCF 10A cells but not in MCF7 cells (panel e) which are reported to be low to negative for its expression. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.