Peter Parker / Spider-Man (voice)
Christopher Daniel Barnes
Mary Jane Watson (voice)
Sara Ballantine
Felicia Hardy (voice)
Jennifer Hale
Aunt May Parker (voice)
Linda Gary
Harry Osborn (voice)
Gary Imhoff
J. Jonah Jameson (voice)
Edward Asner
Joe "Robbie" Robertson (voice)
Rodney Saulsberry
Kingpin (voice)
Roscoe Lee Browne
Michael Morbius (voice) / Richard Fisk (voice) / Morbius (voice)
Nick Jameson
Anna Watson (voice)
Majel Barrett
Alistair Smythe (voice) / Ultimate Slayer Smythe (voice)
Maxwell Caulfield
Norman Osborn (voice) / Norman Osborn / Green Goblin (voice)
Neil Ross
Debra Whitman (voice)
Liz Georges
Flash Thompson (voice)
Patrick Labyorteaux
Detective Terri Lee (voice)
Dawnn Lewis
Landon (voice)
David Warner
Doctor Octopus (voice) / Otto Octavius
Efrem Zimbalist Jr.
Madame Web (voice)
Joan Lee
Newscaster (voice) / News Reporter (voice) / News Announcer (voice)
John Beard
Shocker (voice) / Man-Spider (voice)
Jim Cummings
Beyonder (voice) / Red Skull (voice)
Earl Boen
Eddie Brock / Venom (voice) / Eddie Brock (voice)
Hank Azaria
Hobgoblin (voice) / Jason Phillips / Jason Phillips (voice)
Mark Hamill
Dr. Curt Connors / Lizard (voice)
Joseph Campanella
Rhino (voice)
Don Stark
Kraven (voice) / Quentin Beck / Mysterio (voice)
Gregg Berger
Liz Allen (voice)
Marla Rubinoff
Captain America (voice) / Brace Huntington (voice)
David Hayter
Arthur Avis / Young Vulture (voice) / Vulture (voice)
Alan Johnson
Dr. Mariah Crawford (voice) / Dr. Mariah Crawford / Calypso (voice)
Susan Beaubian
Scorpion (voice)
Richard Moll
Tony Stark / Iron Man (voice)
Robert Hays
Scorpion (voice) / Mac Gargan
Martin Landau
Storm (voice) / Martha Robertson (voice)
Iona Morris
Silvermane (voice)
Jeff Corey
Nick Fury (voice)
Jack Angel