For immunofluorescence analysis, HCT 116 cells were fixed and permeabilized for detection of endogenous CDC27 using Anti-CDC27 Rabbit Polyclonal Antibody (Product # 720461,1:250) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of CDC27 protein (green), Panel b is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating predominant nuclear localization of CDC27 Panel e) represents control cells without primary antibody to assess background. The images were captured at 60X magnification.