- Paul Soles
Peter Parker/Spider-Man
- Peg Dixon
Betty Brant
- Bernard Cowan
Narrator, Dr Matto Magneto, Przywódca Plutonian
- Paul Kligman
J. Jonah Jameson
- Carl Banas
Scorpion, Dr. Banas
- Vernon Chapman
Doctor Octopus
- Ed McNamera
Rhino, Blackbeard, Vulcan
- Gillie Fenwick
Dr. Curt Connors/Lizard, Vulture, Pardo, Plotter, Professor Smarter
- Len Carlson
Green Goblin, Parafino, Stan Patterson/Human Fly, Bolton, Kapitan Ned Stacy
- Tom Harvey
Electro, Sandman, Baron Von Rantenraven, Dr. Atlantean, Dr. Stillwell, Master Vine, Mugs Riley
- Henry Ramer
Dr. Noah Boddy, Grandini the Mystic, Henry Smythe, Lee Patterson/Human Fly
- Billie Mae Richards
Billy Connors
- Chris Wiggins
Mysterio, Blackwell the Magician, Boomer, Harley Clivendon, Infinata