Immunofluorescence analysis of TLR2 was performed using THP-1 cells and THP-1 treated with PMA (10 ng/mL, 20 hours) and LPS (20 ng/mL, 6 hours). The cells were fixed with 4% paraformaldehyde for 10 minutes and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with TLR2 Goat Polyclonal Antibody (Product # PA5-17492) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green) in THP-1 treated cells. 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 merged image of THP-1 treated cells, which shows higher expression for TLR2 protein showing localization in lipid rafts in membrane. Panel e represents the merged image of THP-1 cells, that shows lower expression for TLR2 protein. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.