Alcmene alongside her husband first appears after Pain and Panic feed a baby Hercules a mortality potion, which turned Hercules mortal but left his superhuman strength intact because he did not drink the whole potion. Pain and Panic flee after hearing their voices, and convince Hades that they had eliminated him despite the lie. Meanwhile, Alcmene and her husband decide on raising Hercules themselves after believing that he was abandoned. During this time, they discover a medallion marked with the symbol of the gods on it and decide to keep in a secret until Hercules was old enough to know whom his parents maybe.
Years later, after Hercules has grown into a young lad, he accidentally chaos chaos in the small village he lives in, causing all the villagers to jeer him and his father Amphytyron. Feeling ashamed for his actions, Alcmene and Amphytyron decide to reveal the medallion they hid from Hercules since he was an infant, revealing that he is actually related to the gods and explaining why he has superhuman strength. Hercules decides to set out traveling to the temple of Zeus to find out how he is related to the gods.
Alcmene and Amphytyron do not appear again until after Hercules saves Thebes from the Hydra, where they are shown briefly celebrating there success of their adopted son, which not only made him rich, but also themselves as well.
Alcmene and her husband are then seen again after Hercules saves Megara and Olympus and defeated Hades, where they join Hercules, Meg, Philoctetes, and Pegasus in watching as Zeus constructs a constellation of Hercules and fulfilling Phil's lifelong dream after one of the citizens shouts "That's Phil's boy!".
- In the original tale, Alcmene is actually Hercules' biological mother, whilst her husband Amphytyron is his step-father. In addition, they also have at least two other children in the original tale named Iphicles and Laonome.
- In the film, they are portrayed as peasants within a suburban or rural area of Greece, however in the the original tale they are actually royalty, being the children of two different kings.