Ben Parker first learned this lesson in the late 1940s. Ben's best friend, Keene Marlow was a superhero called the Destroyer. One night Keene and his wife, Lenora Sharp-Marlow were walking down the street when Keene witnessed a robbery. Without hesitation Keene activated his super soldier powers and attacked the thieves. However, when the thieves fired their guns at Keene they accidentally shot and killed Lenora. After this Keene Marlow and Ben Parker learned the lesson, with great power comes great responsibility.
Ben later taught this lesson to his nephew, Peter Parker.
However, Peter did not truly learn the importance of this lesson until after Ben was killed. As Spider-Man, Peter tracked down his uncle's killer he discovered that that it was a thief that he purposely let escape from a wrestling arena. Peter realized that if he had not ignored his responsibility and stopped the thief his Uncle Ben would have still been alive. It was at that moment the Peter truly learned that meaning of with great power comes great responsibility. Peter then began to use his spider powers to help others as Spider-Man.
After Peter was framed by Mysterio, Peter decided to quit being Spider-Man. Peter was later approached by Detective Terri Lee who told him that she believed Spider-Man had been set up and asked for Peter's help in clearing Spider-Man's name. However, Peter refused. Terri Lee then told Peter that Spider-Man never turned his back on anyone. As Terri Lee drove off Peter realized that she was right that Spider-Man never turned his back on anyone except for one time. Peter then remembered back to when he let the burglar escape and his uncle was killed. Peter also remembered the lesson he learned that night that with great power comes great responsibility. Peter then decided to help take down Mysterio as Spider-Man.
When Peter was bonded to the alien symbiote the symbiote started to make him much more violent. So violent that he almost killed Shocker. As Spider-Man was about to kill Shocker, Shocker shouted that he couldn't do this. Spider-Man replied that he has the power and that he could kill him if he wanted. However, Spider-Man remembered what his Uncle Ben always use to tell him, "With great power comes great responsibility." Spider-Man then decided not to kill Shocker. However, the symbiote oozed out from his body and pushed Shocker over a ledge. Spider-Man then quickly caught Shocker with a web line and saved him. At that moment Spider-Man realized the symbiote was corrupting him and he used the sonic vibrations from the church bell to remove the symbiote.
Black Cat also learned this lesson after becoming a superhero.
- Christopher Daniel Barnes felt that Spider-Man's "With great power comes great responsibility" was not as pretentious as Superman's "Truth, justice, and the American way."
- In the comics, the moral saying was not spoken by Uncle Ben and appears in a narrative caption of the comic's last panel, rather than as spoken dialogue.