Katharine Hepburn

Mary Stuart

Florence Eldridge

Elizabeth Tudor

Fredric March

Bothwell

Douglas Walton

Darnley

John Carradine

Rizzio

Robert Barrat

Morton

Gavin Muir

Leicester

Ian Keith

Moray

Moroni Olsen

John Knox

William Stack

Ruthven

Ralph Forbes

Randolph

Alan Mowbray

Throckmorton

Frieda Inescort

Mary Beaton

Donald Crisp

Huntly

David Torrence

Lindsay

Molly Lamont

Mary Livingstone

Anita Colby

Mary Fleming

Jean Fenwick

Mary Seton

Lionel Pape

Burghley

Alec Craig

Donal

Mary Gordon

Nurse

Monte Blue

Messenger

Robert Warwick

Sir Francis Knollys

Halliwell Hobbes

Man (uncredited)

Nigel De Brulier

Judge