| Clark Gregg | Phil Coulson |
| Ming-Na Wen | Melinda May |
| Brett Dalton | Grant Ward |
| Chloe Bennet | Daisy 'Skye' Johnson |
| Iain De Caestecker | Leo Fitz |
| Elizabeth Henstridge | Jemma Simmons |
| Nick Blood | Lance Hunter |
| Adrianne Palicki | Bobbi Morse |
| Henry Simmons | Alphonso 'Mack' Mackenzie |
| Luke Mitchell | Lincoln Campbell |
| Powers Boothe | Gideon Malick |
| Constance Zimmer | Rosalind Price |
| Andrew Howard | Luther Banks |
| Matthew Willig | Lash |
| Spencer Treat Clark | Werner von Strucker |
| Daz Crawford | Kebo |
| Hector Hugo | Jerome Deschamps |
| Blair Underwood | Andrew Garner |
| Marco Martinez | Officer Stuart
(as Marco Antonio Martinez) |
| Anthony Corrales | S.H.I.E.L.D. Agent |
| Mike Miller | S.H.I.E.L.D. Medic |
| Nick Brett | Hydra Arsonist
(uncredited) |