Josh Keaton
Peter Parker/Spider-Man (voice)
Lacey Chabert
Gwen Stacy (voice)
James Arnold Taylor
Harry Osborn (voice)
Alan Rachins
Norman Osborn (voice)
Deborah Strang
Aunt May Parker (voice)
Ben Diskin
Eddie Brock (voice)
Dee Bradley Baker
Curt Connors/Lizard (voice)
Robert Englund
Adrian Toomes/Vulture (voice)
Crispin Freeman
Max Dillon/Electro (voice)
Jeff Bennett
Montana/Shocker (voice)
Daran Norris
J. Jonah Jameson (voice)
Joshua LeBar
Flash Thompson (voice)
Peter MacNicol
Otto Octavius/Doctor Octopus (voice)
Alanna Ubach
Liz Allen (voice)
Grey DeLisle
Sally Avril (voice), Betty Brant (voice)
Clancy Brown
Alex O'Hirn/Rhino (voice)