For immunofluorescence analysis, SHSY-5Y cells were fixed and permeabilized detection of endogenous DAPK1 using Anti-DAPK1 Recombinant Rabbit Monoclonal Antibody (Product # 703639, 1:250) and labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Panel a-d) clearly demonstares cytoplasmic localization of DAPK1 in SHSY-5Y cells . Panel e-h) shows no specific staining in MCF7 cells which is DAPK1 negative,demonstrating specificity. The images were captured at 60X magnification.