Though this takes place in one of She-Hulk's comics, Spidey's response◊ to why Jameson hates him so much.
Now doubles as Hilarious in Hindsight, considering what happened over in the Ultimate 'verse.
Also, JJJ's use of the classic defense, "I have black friends", is actually true, considering his buddy Joe Robertson. This makes it even funnier (along with his lawyer's Face Palm).
Lady Stilt-Man. Even Spidey can't stop laughing... And the way she is defeated is even more hilarious. Even before Deadpool shows up, this is one of the most hilarious issues of Spider-Man in years.
2003's "The Rules of the Game" arc is packed with them. A robot falling in love with Reed Richards, Rocket Racer smacking into a window in an attempt to make a cool entrance, Peter putting on a disguise and getting beaten up at a Bad Guy Bar... but the best had to be Scorpion being tricked into into breaking his own leg. "If that was... then whose leg is... Oh."
One issue of Spider-Man was called "Slyde Into Destiny". It features Spidey's unknown archfoe, an incredibly minor and deluded villain calling himself "Slyde".
Slyde: That's right, citizens of New Slyde City! You have all been Slyde'd! Slode? Slydomized?
During Mike Wieringo's Spider-Man run, just after he super-evolved and just before the Civil War. Aunt May completely owns, humiliates and scares the living bejeebers out of Tony Stark, Reed Richards and Hank Pym. These three have faced down cosmic horrors from beyond time and space and one little old lady turns them into quivering messes.
Aunt May has frightened J. Jonah Jameson so badly that JJJ has forbidden Peter from ever letting her phone him again. This is a (perfectly normal) man whose reaction to costumed killers tearing his office apart to murder him is one of irritation at how much this is going to cost, and one little old lady had him holding the phone two feet from his ear and cringing. Do not mess with Aunt May.
Batman / Spider-Man, after Batman corrects Spidey's reference to The Road To Morocco:
Spider-Man: A-ha! I knew we had something in common besides our love of tights! You're into old movies too, huh?
Batman: I was - when I was young.
Spider-Man: Young? Somehow, I can't picture you ever being young! You must've been pretty cute, waddling around in your diapers, chasing the Joker across the playground!
Batman: I was never "cute".
In one occasion the FF had to Spot the Imposter between Spider-Man and the Chameleon. Ben solved it by throwing both of them down of the building, with poor Chameleon being recognized when he started screaming while Spidey just spun a web and flung himself away.
The two-parter "Hyde and Seek" is full of humorous moments, such as Spidey suspecting that danger's "right around the corner", only to find that Hyde has thrown an entire corner of a building at him. Or Spider-Man showing off a defeated villain to someone saying "Daddy, Daddy! Can I keep him?"
Amazing Spider-Man #699, as Peter Parker, trapped in Dr. Octopus' body, attempts to figure out how Ock was able to swap bodies with him, starts to look through Ock's memories... and finds out he and Aunt May did the nasty!
Peter:[Mentally] NO! NOOOOO! [Vocally, as Doc Ock] ...Ahhhhhh...!
Peter Parker: Spider-Man wasn't trying to attack the city, he was trying to save it. That's slander. J. Jonah Jameson: It is not! I resent that! [beat] Slander is spoken. In print, it's libel.
The scene where Flash tries attacking Peter after he's gotten his powers. His super-reflexes are represented by Bullet Time... which is so slow that he has time to dodge Flash's punch, shoot a confused look up at Flash, then focus his attention back on the fist, all in the time it takes a single punch to be thrown.
Spider-Man silencing J. Jonah Jameson with webbing and telling him (referring to him and Green Goblin) "Hey kiddo, let Mom and Dad talk for a minute will ya?"
Peter's original idea for his name.
Announcer: The Human Spider, that's it, that's the best you got?
Peter: Yeah.
Announcer: Oh, that sucks...
Peter to Bonesaw: "That's a cute outfit. Did your husband give it to you?"
Even before the match, there's a funny moment with Peter when he initially wants to sign up, courtesy of Octavia Spencer herself: "There's no featherweight division here, small fry. Next!"