- Prologue/Epilogue
- Henry VIII – King of England
- Cardinal Wolsey – Archbishop of York and Lord Chancellor; initially, Henry's chief adviser
- Queen Katherine – later divorced
- Anne Bullen – Katherine's maid of honour; later Queen Anne[a]
- Duke of Buckingham – hates Wolsey, who charges him with treason
- Thomas Cranmer – Archbishop of Canterbury
- Stephen Gardiner – close ally of Wolsey; King's secretary; later Bishop of Winchester
- Lord Chamberlain – historically, the play covers a period in which the position was held by both Somerset and Lord Sands, but the play presents the character as one consistent figure[5]
- Duke of Norfolk – Anne Bullen's grandfather; later Earl Marshal
- Duke of Suffolk – Henry's brother-in-law; later High Steward
- Earl of Surrey – Buckingham's son-in-law and Norfolk's son; also Lord Lieutenant of Ireland
- Cardinal Campeius – Papal legate sent to judge legitimacy of Henry's marriage to Katherine
- Lord Caputius – ambassador of Charles V, Holy Roman Emperor (Katherine's nephew)[b]
- Sir Thomas Cromwell – Wolsey's secretary and protégé; later secretary to the Privy Council
- Lord Sands – historically, was Lord Chamberlain during some of the period covered by the play[5]
- Lord Abergavenny – Buckingham's son-in-law
- Lord Chancellor (Thomas More) – More replaces Wolsey, after which time he is never mentioned by name.[7] Thomas Audley, 1st Baron Audley of Walden replaced More.
- Bishop of Lincoln
- Sir Thomas Lovell – Chancellor of the Exchequer
- Sir Henry Guildford – Master of the Horse
- Sir Nicholas Vaux – Governor of Guînes
- Sir Anthony Denny – Groom of the Stool
- Dr. Butts – Henry's personal physician
- Griffith – Katherine's usher
- Garter King-of-Arms
- Buckingham's Surveyor – historically, Charles Knevet[5]
- Brandon – an officer[c]
- Serjeant-at-Arms
- Porter
- Old Lady – Bullen's chaperone
- Patience – Katherine's waiting woman
- Doorkeeper of the Council Chamber
- First Secretary – works for Wolsey
- Second Secretary – works for Wolsey
- First Scribe – at Katherine's trial
- Second Scribe – at Katherine's trial
- Crier
- Gardiner's Page
- Three Gentlemen
- Lord Chamberlain's Servant
- Porter's Man
- Messenger
- At the Legatine Court
- Archbishop of Canterbury – predecessor of Thomas Cranmer (non-speaking role)
- Bishop of Ely (non-speaking role)
- Bishop of Rochester (non-speaking role)
- Bishop of Saint Asaph (non-speaking role)
- At Bullen's coronation
- Marquis Dorset (non-speaking role)
- Bishop of London (non-speaking role)
- Lord Mayor of London — historically Sir Stephen Peacock[10] (non-speaking role)
- At Elizabeth's christening
- Infant Princess Elizabeth (non-speaking role)
- Marchioness Dorset – Elizabeth's godmother (non-speaking role)
- Duchess of Norfolk – Elizabeth's godmother (non-speaking role)
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