Immunofluorescent analysis of the neurofilament medium chain in paraffin-embedded human brain tissue (right) compared to a negative control without primary antibody (left). Tissue sections were deparaffinized with xylene, and rehydrated with ethanol. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and microwaved for 8-15 min. Following antigen retrieval, tissues were washed with water and PBS, and then blocked in 0.3% BSA for 30 min at room temperature. Tissues were then probed with a neurofilament light chain monoclonal antibody in 0.3% BSA at a dilution of 1:50 for 1 hour at 37°C. Tissues were then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 488 conjugate for 1 hour at 37°C (green). Nuclei (blue) were stained with DAPI. Images were taken at 40X magnification.