Acanthamoeba (AGE isolate belonging to T1 genotype) proteases exhibit optimal activities at various temperatures. To determine the effects of variable temperatures, zymographic assays were performed as described in the "Materials and Methods" and gels incubated at various temperatures. Note that 37 – 50°C produced optimal proteolytic activities. Results are representative of three independent experiments. Each lane contained 0.5 μg protein.