The title of this episode is taken from the phrase, "Along came a spider...", which is a line from the nursery rhyme, "Little Miss Muffet". The complete rhyme is as follows:
Little Miss Muffet
Sat on a tuffet,
Eating of curds and whey;
Along came a spider
Who sat down beside her
And frightened Miss Muffet away.
This episode provides the origin of Spider-Man. Spidey's origin was originally chronicled in his first appearance, which was in Amazing Fantasy #15 in 1962. The origin provided here remains close to the canon history with Peter Parker being bitten by a radioactive spider, thus gaining his powers and learning the hard-valued lesson of power and responsibility following the death of his Uncle Ben.
A billboard seen at the beginning of the episode promotes a newspaper article called "Who Is Spider-Man?" by Stan Lee of the Daily Bugle. Stan Lee is actually the co-creator of Spider-Man, whereas the Daily Bugle is a fictional newspaper from the comics.
In future issues of the comic, the science lab where Peter is bitten by the spider is part of Osborn Industries, owned by Norman Osborn. Norman Osborn is also Spider-Man's nemesis, the Green Goblin.
Shocker's real name is Herman Schultz. A recurring foe of Spider-Man, he first appeared in Amazing Spider-Man #46 in 1967.