Holloway was born in Neath, Wales, and studied in London at the Guildhall School of Music and Drama in London. After initial engagements, including at the Academy of St. Martin in the Fields and at the English Chamber Orchestra, he became manager and concertmaster of the Kent Opera Orchestra in the 1970s. After an encounter with Sigiswald Kuijken in 1972, he started playing the Baroque violin and gained a reputation as violinist, teacher and conductor in the field of historically informed performance.
In 1970 he became the concertmaster of Sir Roger Norrington's London Classical Players, and later Andrew Parrott's Taverner Consort and Players. Besides playing in numerous Baroque orchestras, he is a noted musicologist and lecturer.
Holloway has taught at the Guildhall School of Music and Drama in London, the Schola Cantorum in Basel, and the Early Music Institute of Indiana University in Bloomington. He has given classes and led workshops in most European countries, as well as in Korea, New Zealand and the USA. In 2004, he was Regents’ Lecturer at UC Berkeley. In 1999, he began to teach at the Hochschule für Musik "Carl Maria von Weber" in Dresden.
Between 2003 and 2005 Holloway served as musical director of the Indianapolis Baroque Orchestra, and in 2005 and 2006 concertmaster and music director of the period instrument ensemble and orchestra known as New Trinity Baroque.
In 2005 he founded jointly with Belgian conductor and harpsichordist Florian Heyerick and a music agent the Mannheimer Hofkapelle, which in the summer of 2007 could be heard for the first time in 300 years with its original complement of 40 musicians. Between 2006 and 2012, he was artistic director of the international violin competition and master class known as Violine in Dresden.
Holloway has appeared on more than 100 CD recordings, and is the winner of a Gramophone Award in 1991 for his recording of Biber's Mystery Sonatas. He won two Danish Grammy Awards for his recordings of the chamber music and vocal music of Dieterich Buxtehude (1994 and 1997). His CD recordings of the Rosary Sonatas by Heinrich Ignaz Biber and of Sonatas Opus 5 of Jean-Marie Leclair won the Preis der Deutschen Schallplattenkritik ("German Record Critics' Award"). He has recorded Bach's sonatas and partitas for solo violin.. In addition to recordings of The Four Seasons and L'estro Armonico by Antonio Vivaldi, he recorded the complete chamber works of Georg Friedrich Händel with the ensemble L'Ecole d'Orphée he founded in 1975.
- von Rhein, John (18 February 1993). "Holloway and friends do 17th Century works proud". Chicago Tribune. p. 28.
- Ruhe, Pierre (12 November 2005). "He seems to like being a pioneer: Period violin eminence John Holloway on taking up with Atlanta's New Trinity Baroque". The Atlanta Journal-Constitution. Archived from the original on 19 July 2009. CS1 maint: discouraged parameter (link)
- "Archived copy". Archived from the original on 5 July 2010. Retrieved 7 May 2010. CS1 maint: discouraged parameter (link) CS1 maint: archived copy as title (link) Artist profile: John Holloway, Aloysia Assenbaum and Lars-Ulrik Mortensen