Immunofluorescent analysis of PPAR gamma (red) in 3T3-L1 differentiated day 7 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a PPAR gamma rabbit polyclonal antibody (Product # PA1-824) at a dilution of 1:200 in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit IgG (H+L) Secondary Antibody, Dylight 680 (Product # 35568) at a dilution of 1:1000 for at least 30 minutes at room temperature in the dark (red). Lipids (green) were stained with HCS LipidTOX neutral lipid stain (Product # H34475) at a dilution of 1:200 for at least 30 minutes at room temperature in the dark. Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a EVOS FL Auto Imaging System at 20X magnification.