The activity and distribution of the lysosomal marker enzyme acid phosphatase have been studied in the epidermis during tail regeneration in the lizard Podarcis muralis. This study gives information both on the mechanism of separation of the old from the new epidermal generation and on the involvement of lysosomes in β- and α-keratinization in lepidosaurians. The biochemical analysis shows that the activity of acid phosphatase was higher in the regenerating tail than in the normal tail until the stage of skin shedding at around 25–30 days post-amputation. The enzyme was present in keratinizing layers of epidermis, and particularly in the shedding layer. The shedding layer followed the outline of the regenerating scales beneath the wound in the epidermis showing that lysosomal enzymes are involved in epidermal shedding. The ultrastructural study showed that acid phosphatase was initially localized within the lysosomes but, with the progress of differentiation, it also appeared free within the cytoplasm of the cornifying cells of the shedding complex and of the β-keratinizing cells, and in the intercellular space among them. The role of lysosomal enzymes in the shedding process and their involvement in the mechanism of β-keratinization is discussed. While the process of β-keratinization has an intracellular and extracellular localization of acid phosphatase, the process of α-keratinization shows that most acid phosphatase is localized within the cell. Lysosomes and mesos granules containing acid phosphatase were concentrated in the cytoplasm of differentiating mesos cells. Some mesos granules later discharged their contents into the extracellular space. The α-layer was partially formed in proximal regenerated scales around 30 days post-amputation. Lysosomes appeared to concentrate within pre-keratinizing cells of the α-keratin layer and little reactivity was seen extracellularly.