Peter Parker / Spider-Man (voice)

Christopher Daniel Barnes

J. Jonah Jameson

Edward Asner

Mary Jane Watson (voice)

Sara Ballantine

Felicia Hardy (voice)

Jennifer Hale

Aunt May Reilly-Parker

Linda Gary

Joseph `Robbie' Robertson

Rodney Saulsberry

Harry Osborn (voice)

Gary Imhoff

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