Anubis serves as the main antagonist in the animated film Yu-Gi-Oh! The Movie: Pyramid of Light. According to the movie's novelization, he is an evil sorcerer who co-created the Pyramid of Light alongside a man named Aknadin. This artifact was intended as a failed replica of the Millennium Puzzle, but Anubis stole it for his own purposes, seeking to use its power to destroy all life on Earth. His initial reign of terror was stopped millennia ago by the Pharaoh Atem, who used the Dagger of Fate to defeat him. Though seemingly vanquished, Anubis managed to seal his own soul inside the Pyramid of Light, allowing him to await the chance to return.

Physically, Anubis appears in several forms. When he first reveals his incomplete human-like form, he is a tall, tanned, and muscular man with pulsating skin, dirty blonde hair, and red eyes, with a jewel embedded in his forehead. He wears a ragged black cape, spiked padding, gray shoulder guards, and the Pyramid of Light itself hangs around his neck. In a vision experienced by Yugi Muto, Anubis is seen with the head of a black jackal with a white muzzle. After he is defeated in a Duel, he uses the power of the Pyramid of Light to assume a final monstrous shape: an armored, bipedal, jackal-like creature with bladed wings and an axe-like blade atop his head, with the pyramid's jewel embedded in his chest.

In terms of personality, Anubis is a ruthless, power-hungry, and vengeful sorcerer, a stark contrast to the benevolent god of death from Egyptian mythology with whom he shares a name. He is consumed by a desire to eradicate humanity, viewing civilization as weak and corrupt. He is highly cunning and manipulative, preferring to operate from the shadows. Rather than confronting his enemies directly at first, he subtly orchestrates events, such as slipping the Pyramid of Light card into Maximillion Pegasus's collection to ensure it would eventually fall into Seto Kaiba's hands. Anubis is also deeply arrogant, confident in his superiority over mortals and even the Pharaoh, a flaw that ultimately contributes to his downfall. He taunts his opponents and revels in their suffering, refusing to accept defeat even when his plans begin to crumble.

Anubis's role in the story is that of the primary instigator. After being sealed for five thousand years, his spirit is reawakened when Yugi Muto solves the Millennium Puzzle. His physical remains and the Pyramid of Light are eventually moved to a museum in Domino City, allowing his spirit to roam free. He proceeds to manipulate Kaiba into activating the Pyramid of Light card during a Duel against Yugi, triggering a Shadow Game. Anubis then takes over the Duel himself, summoning his Sphinx monsters. He also invades the inner world of the Millennium Puzzle, trapping Yugi and his friends and draining the life force of the Pharaoh Atem to restore his own body. His key relationships are defined by his ancient rivalry with Pharaoh Atem and his role as a manipulator of Seto Kaiba, whose competitive nature he exploits as a tool to achieve his own resurrection.

Throughout the film, Anubis shows little in the way of character development, remaining a single-mindedly destructive force. However, his story arc follows a clear trajectory from a sealed, spiritual entity to a fully resurrected physical threat. He begins as an invisible schemer, then becomes an active participant in the Duel, and finally transforms into a massive, real-world monster when his initial plan fails. His most notable abilities are almost entirely derived from the Pyramid of Light. This artifact grants him a wide range of powers, including the ability to travel spiritually, command an army of undead servants, influence the thoughts of others, and show people visions of the future. When the Pyramid of Light card is activated in a Duel, Anubis can drain the life energy of anyone who takes damage, using it to heal himself. He can also create cards out of thin air and materialize his Sphinx monsters directly from a black slime. In his final monster form, he can breathe a devastating black flame and is immensely resilient. The presence of his true form also has the power to make the monsters and effects of Duel Monsters cards become fully real. He primarily uses a Sphinx-themed Deck, focusing on summoning Andro Sphinx, Sphinx Teleia, and their combined form, Theinen the Great Sphinx, whose attack points he can raise to overwhelming levels. Despite his vast power, he is ultimately vulnerable to the Blue-Eyes Shining Dragon's Shining Nova ability, which sacrifices itself to destroy him and the Pyramid of Light once and for all.