6-AF488 tyramide is a bright, green fluorescent dye (Ex=496 nm, Em=524 nm). It serves as a fluorescent substrate for horseradish peroxidase (HRP)-catalyzed deposition, making it useful for tyramide signal amplification (TSA). Furthermore, 6-AF488 tyramide can be utilized in multiplex immunohistochemistry (mIHC) experiments.

6-AF488 tyramide detection involves several precise steps. First, place tissue sections in each well of a 24-well plate, ensuring they remain moist. Then permeabilize the tissue with PBS containing 0.2% Triton X-100 for 5-10 minutes. Wash with PBS-T for 5-10 minutes, followed by incubation in a 1% H₂O₂ solution for 20 minutes to block endogenous peroxidase activity. Rinse three times with PBS for 5-10 minutes each, and incubate the tissue for 1 hour in PBS with 1% blocking solution (4% normal donkey serum + PBS-T) to prevent nonspecific binding. Incubate with primary antibody at room temperature for 2 hours or overnight at 4°C, then wash with PBS three times. Incubate with HRP-conjugated goat anti-rabbit secondary antibody (1:100 in 1% blocking solution) at room temperature for 2 hours, followed by three PBS washes. Activate TSA buffer with 0.0015% H₂O₂, incubate the tissue in 6-AF488 tyramide working solution (1:100 in activated TSA buffer) for 5-10 minutes. Wash with PBS three times, mount sections on slides, position slides vertically in a rack for 10 minutes to dry, then rinse quickly with ddH₂O. Use AntiFade Mounting Medium (with DAPI) for mounting. Finally, examine with a fluorescence microscope. Note: Adjust the concentration of the 6-AF488 tyramide working solution as needed.