| Tobey Maguire | Peter Parker / Spider-Man |
| Kirsten Dunst | Mary Jane Watson |
| James Franco | Harry Osborn / New Goblin |
| Thomas Haden Church | Flint Marko / Sandman |
| Topher Grace | Eddie Brock / Venom |
| Bryce Dallas Howard | Gwen Stacy |
| Dylan Baker | Curt Connors |
| Rosemary Harris | May Parker |
| J.K. Simmons | J. Jonah Jameson |
| James Cromwell | Captain Stacey |
| Theresa Russell | Mrs. Marko |
| Bill Nunn | Joseph Robertson |
| Bruce Campbell | Maître D' |
| Elizabeth Banks | Miss Brant |
| Ted Raimi | Hoffman |
| Perla Haney-Jardine | Penny Marko |
| Willem Dafoe | Norman Osborn / Green Goblin |
| Cliff Robertson | Ben Parker |
| Elya Baskin | Mr. Ditkovitch |
| Mageina Tovah | Ursula |
| John Paxton | Houseman |
| Becky Ann Baker | Mrs. Stacy |
| Stan Lee | Man in Times Square |
| Michael Papajohn | Dennis Carradine / Carjacker |
| Joe Manganiello | Flash Thompson |
| Hal Fishman | Anchorman |
| Lucy Gordon | Jennifer Dugan |
| Steve Valentine | Photographer |
| Tim Maculan | Play Director |
| Marc Vann | Play Producer |
| Joe Bays | Jazz Club Manager |
| Gregg Daniel | Precinct Detective |
| Rogelio T. Ramos | Emergency Room Doctor |
| Timothy Patrick Quill | Crane Operator |
| Christopher Young | Pianist |
| Grant Curtis | Driver |
| Toni Wynne | Congratulatory Woman at Daily Bugle |
| Andre B. Blake | Crane Disaster Radio Policeman |
| Kathryn Bryding | Woman Outside Theater |
| Joe Virzi | Police Detective |
| Bill E. Rogers | New Jersey State Policeman |
| Michael Alexander | New Jersey State Policeman |
| April Parker-Jones | Test Site Technician |
| Edward Padilla | Test Site Technician |
| Robert Curtis Brown | Test Site Technician |
| Paul Terrell Clayton | Test Site Technician |
| Carolyn Neff | Test Site Technician |
| Christina Cindrich | Test Site Technician |
| Sonya Maddox | ICU Nurse |
| Derrick 'Phoenix' Thomas | Cop at Crane Disaster |
| Jessi Collins | Mary Jane's Replacement |
| Michael McLaughlin | Boy at Key to the City Ceremony |
| Anne Gartlan | Councilwoman |
| Emilio Rivera | Policeman at Sand Truck |
| Keith Woulard | Policeman at Sand Truck |
| Reynaldo Gallegos | Armored Car Driver |
| Jim Coope | Newstand Patron |
| Dean Edwards | Newstand Patron |
| Margaret Laney | Newstand Patron |
| Aimee Miles | Coffee Shop Waitress |
| Tanya Bond | Jazz Club Waitress |
| Mark Kubr | Jazz Club Bouncer |
| Tony Besson | Photoshoot Client |
| Ramon Adams | Businessman (uncredited) |
| A.J. Adelman | Theater Attendee (uncredited) |
| Angelis Alexandris | Pedestrian (uncredited) |
| Dawn Marie Anderson | I.C.U Nurse (uncredited) |
| Frank Anello | NYPD Officer (uncredited) |
| Anya Avaeva | Model (uncredited) |
| David Backus | Bucket Boy (uncredited) |
| Tiffany L. Baker | News Reporter (uncredited) |
| Sujeilee Candele | Rescued Girl (uncredited) |
| Michael Ciesla | Photographer (uncredited) |
| Irina Costa | Jazz Club Waitress (uncredited) |
| John Crann | Theater Patron (uncredited) |
| Crystal Marie Denha | Jazz Club Girl (uncredited) |
| Amy V. Dewhurst | Columbia Grad Student (uncredited) |
| Paul Edney | New York City Commuter / Driver (uncredited) |
| Natalie Fabry | Girl at Key Ceremony (uncredited) |
| Keith Fausnaught | Pedestrian (uncredited) |
| Shaun Patrick Flynn | Businessman (uncredited) |
| Logan Fry | Automobile Driver (uncredited) |
| Kevin Fung | Spectator (uncredited) |
| Tony Galtieri | News Spectator (uncredited) |
| Chuck Gerena | Funeral Limo Driver (uncredited) |
| Brian Hopson | News Reporter (uncredited) |
| Claude Jay | Cop (uncredited) |
| Julie Jei | Female New Yorker (uncredited) |
| Andrew James Jones | Jazz Club Bouncer (uncredited) |
| Natalie Jones | Girl at Key Ceremony (uncredited) |
| Christopher Jude | Student (uncredited) |
| Brittany Krall | ER Nurse (uncredited) |
| Alyssa Lakota | Dog Walker (uncredited) |
| Pierangeli Llinas | Woman in Court (uncredited) |
| Bernadette Lords | British Publicist (uncredited) |
| Sandrine Marlier | Shocked Pedestrian (uncredited) |
| Laura McDavid | Journalist (uncredited) |
| Natalie McNeil | Girl Screaming in Camaro (uncredited) |
| Daniel Mignault | Kid in Times Square (uncredited) |
| Martha Millan | Oscorp Receptionist (uncredited) |
| Michele-Nanette Miller | Girl in Times Square (uncredited) |
| Claudia Katz Minnick | Robbie's Assistant (uncredited) |
| Robert Myers | Theatergoer (uncredited) |
| Jen Oda | Beautiful Girl (uncredited) |
| Anjelia Pelay | Beautiful Woman (uncredited) |
| Nick Poltoranin | Police Officer (uncredited) |
| Vanessa Reseland | Beautiful Woman (uncredited) |
| La Rivers | Beautiful Woman (uncredited) |
| Bria Roberts | Businesswoman (uncredited) |
| Vanessa Ross | Sandman Victim (uncredited) |
| Brenna Roth | Bad Girl (uncredited) |
| Shade Rupe | City Hall Cheerer (uncredited) |
| Arick Salmea | Guy with Ticket (uncredited) |
| Eric Shackelford | Jazz Club Beatnik (uncredited) |
| Daniel Shafer | Jazz Club Guest (uncredited) |
| Abbey Skinner | Café Girl (uncredited) |
| Kristin Somo | Jazz Club Patron (uncredited) |
| Jennifer Sparks | Jazz Club Waitress (uncredited) |
| Christopher Stadulis | Firefighter (uncredited) |
| Arne Starr | Broadway Audience Member (uncredited) |
| Liam Stone | Jazz Club Musician (uncredited) |
| Tajna Tanovic | Girl in Cab (uncredited) |
| Aija Terauda | Model (uncredited) |
| Brigid Turner | Hot Girl (uncredited) |
| Evelyn Vaccaro | Beautiful Woman (uncredited) |
| Nick Vlassopoulos | Bar Patron #5 (uncredited) |
| Sincerely A. Ward | Model (uncredited) |
| Silq Webster | Model (uncredited) |
| Graig F. Weich | Student (uncredited) |
| Fredrick Weiss | Restaurant Patron (uncredited) |
| Jennifer Weston | ER Nurse (uncredited) |
| Trenton Willey | City Hall Cheerer (uncredited) |
| Ray Wineteer | Spectator (uncredited) |
| Emily Eckes | Beautiful Woman (uncredited) |
| Jimmy Star | Police Officer (uncredited) |