His first on-screen role was as a military policeman in the 1952 movie Fearless Fagan. In 1953, he began appearing on television. In 1958, he was cast in the episode "The Trial" of the American Civil War drama Gray Ghost with Tod Andrews. Eisley was in the 1959 Roger Corman film The Wasp Woman, which he described as "a hell of a lot of fun". Eisley's big break was being discovered playing opposite Jerry Paris in a Pasadena, California production of Who Was That Lady I Saw You With? where he was signed to a contract with Warner Bros. In the days of Tab, Ty, and Rock, Warner Bros. did not want a leading man with the name of "Fred" so they changed his first name to "Anthony". In 1959, Eisley played Carter Henry (as Fred Eisley), The Young Philadelphians (1959).

In 1957, Eisley played Joe Foss in the episode "Jose Foss, Devilbird" of the military television series Navy Log. Foss was a World War II Medal of Honor winner who later became the governor of South Dakota and an American sportsman. The episode also features Mason Alan Dinehart as Marly.

Eisley was best known for his starring role in the series Hawaiian Eye, which aired from 1959 to 1963. After being seen in the play Who Was That Lady? he was signed to Warner Bros. Eisley changed his name from Fred to Anthony at the request of the studio. Anthony had thought he would play a Comedian but Warner Brothers Television placed him in a suave private eye role in Hawaiian Eye. Eisley said that he left after the third season to be replaced by Troy Donahue as a hotel social Director, Philip Barton. In the two previous seasons, Donahue had portrayed the detective Sandy Winfield, II, on another ABC/WB series, Surfside 6, set on a houseboat in Miami Beach. Donahue was eleven years Eisley's junior.

During his Warner Brothers period, Eisley also appeared in Portrait of a Mobster (1961).

His most memorable role in film was as Griff in The Naked Kiss (1964), Sam Fuller's controversial attack on alleged American small town hypocrisy. Eisley became known as a cult schlock star for his appearances in Antonio Margheriti's Lightning Bolt (1965), Jack Broder's The Navy vs. the Night Monsters (1966), Journey to the Center of Time (1967), The Mighty Gorga (1969), Al Adamson's Dracula vs. Frankenstein (1971), Ted V. Mikels's The Doll Squad (1974), Monstroid (1980) and Deep Space (1988).

Eisley co-starred as character Clint Braden, suitor to the Nancy Kovack character of Nellie Bly, in the 1966 Elvis Presley vehicle, Frankie and Johnny. He also played Ben Mitchell in the 1968 musical film Star!, starring Julie Andrews.

Easley guest-starred in an episode of the ABC religion drama series Going My Way, with Gene Kelly. He appeared six times in Jack Webb's 1967-1970 revival of Dragnet; in one segment he played a corrupt policeman and once he played an attempted murderer. During the eight-year run of ABC's The F.B.I., with Efrem Zimbalist, Jr., Eisley made 17 appearances as an agent. He also appeared three times on CBS's Perry Mason during the final three seasons of that series. In his second guest appearance in 1964 he played murder victim Vince Rome in "The Case of the Missing Button."

Eisley was a guest as a villain in an episode of The Wild Wild West, with his former Hawaiian Eye co-star Robert Conrad. In 1970, he guest-starred in an episode of The Silent Force.

In 1973, he appeared as Ross Nelson, newscaster at Mary's competitor, channel 8, in the Season 4 episode "WJM Tries Harder" on The Mary Tyler Moore Show. He had appeared with Mary Tyler Moore years before in "The Lady and the Tiger and the Lawyer," a 1964 episode of The Dick Van Dyke Show.

Anthony Eisley died of heart failure 10 days after his 78th birthday, on January 29, 2003 in Woodland Hills, California, and is interred at Forest Lawn Memorial Park (Hollywood Hills).