Katharine Hepburn

Character: Mary Stuart

Florence Eldridge

Character: Elizabeth Tudor

Fredric March

Character: Bothwell

Douglas Walton

Character: Darnley

John Carradine

Character: Rizzio

Robert Barrat

Character: Morton

Gavin Muir

Character: Leicester

Ian Keith

Character: Moray

Moroni Olsen

Character: John Knox

William Stack

Character: Ruthven

Ralph Forbes

Character: Randolph

Alan Mowbray

Character: Throckmorton

Frieda Inescort

Character: Mary Beaton

Donald Crisp

Character: Huntly

David Torrence

Character: Lindsay

Molly Lamont

Character: Mary Livingstone

Anita Colby

Character: Mary Fleming

Jean Fenwick

Character: Mary Seton

Lionel Pape

Character: Burghley

Alec Craig

Character: Donal

Mary Gordon

Character: Nurse

Monte Blue

Character: Messenger

Robert Warwick

Character: Sir Francis Knollys

Halliwell Hobbes

Character: Man (uncredited)

Nigel De Brulier

Character: Judge