Holloway made his motion picture debut in 1921, appearing in a silent movie engagingly entitled The Rotters. That same year, working in collaboration with fellow singing comedian Leslie Henson, Holloway helped to form a London-based touring musical revue called the Co-Optimists. This troupe would virtually reinvent itself every year through 1929, and they were filmed in 1930. Holloway established himself as a BBC radio personality in 1923 and developed his solo act throughout the 1920s while continuing his involvement with the musical theater. He was cast as Bill Smith in the London production of Vincent Youmans' musical comedy Hit the Deck (1927), in Song of the Sea (1928), and in Coo-ee (1929). He began recording his monologues for the phonograph market in the early '30s. Peopled with colorful "North Country" characters named Albert, Sam Small, and the Ramsbottoms, these remarkable recitations were couched in rhyming stanzas carefully crafted by George Marriott Edgar, Greatrex Newman, Robert Patrick Weston, and Bert Lee, and also by Holloway himself, who occasionally sang the verses rather than declaiming them.

Holloway participated in a number of stage revues during the 1930s. These included The Savoy Follies (1932), Three Sisters (1934), Here We Are Again (1935), All Wave (1936), London Rhapsody (1938), Up and Doing (1940), and Fine and Dandy (1942). He showed up in a steady stream of British movies including Sing as We Go (1934); Squibs (1935); The Vicar of Bray (1936); a film adaptation of George Bernard Shaw's Major Barbara (1941); Champagne Charlie, The Way Ahead, and This Happy Breed (1944); Caesar and Cleopatra, Brief Encounter, and The Way to the Stars (1945); Meet Me at Dawn (1946); Nicholas Nickelby (1947); and Another Shore, The Winslow Boy, and Hamlet, wherein he played the Gravedigger opposite Sir Lawrence Olivier (1948). He appeared in the films The Perfect Woman and Passport to Pimlico (1949); The Magic Box and The Lavender Hill Mob (1951); The Titfield Thunderbolt and The Beggars Opera (1952); Meet Mr. Lucifer (1953); An Alligator Named Daisy (1955); No Tree in the Street and Hello, London (1959); and No Love for Johnnie (1961). On-stage in New York he played Bottom in Shakespeare's A Midsummer Night's Dream and personified Alfred P. Doolittle in both the stage (1956, 1958) and screen (1964) versions of My Fair Lady.

In 1957 Holloway made some of his all-time funniest and best recordings in the company of Beatrice Lillie and Cyril Ritchard (Nonsense Verse of Carroll and Lear, Cadmon Literary Series LP TC 1078). His dramatic readings of "The Pig-Tale" from Lewis Carroll's Sylvie and Bruno and Edward Lear's heart-rending existential opus The Dong with the Luminous Nose were unparalleled triumphs of theatrical timing, artful delivery, and seasoned wit. In 1960 Stanley Holloway became an officer of The Most Excellent Order of the British Empire, and toured as a solo act under the billing Laughs and Other Events. In 1962 he was cast as an English butler in the U.S. television sitcom Our Man Higgins, while 1964 found him on-stage in Philadelphia in Cool Off!, a Faustian spoof. Wiv a Little Bit o Luck: The Life Story of Stanley Holloway as told to Dick Richards was published in 1967 and reprinted in 1981. Holloway toured Asia and Australia in 1977 with The Pleasure of His Company, a Noël Coward tribute show, gave a Royal Command Performance in 1980, and died at the age of 91 in a nursing home in Littlehampton (Sussex) England on January 30, 1982.