Unlike similar programs such as The Ed Sullivan Show, the series used a different host each week. Among the performers and hosts on the show were Bing Crosby (who made the first and the most appearances as guest host: 31 in all, including his family on several of the annual Christmas shows), Dean Martin, Liberace, Frank Sinatra, Milton Berle, Sammy Davis Jr., Sid Caesar, Peter Lawford, The Rolling Stones, Groucho Marx, Joan Crawford, Bette Davis, Tony Bennett, Judy Garland, Jimmy Durante, The Supremes, Ginger Rogers, The Temptations, Dusty Springfield, Phyllis Diller, Elizabeth Montgomery, and many others. (Martin's hosting was a bit ironic, given that the slot had been vacated by his ex-partner Jerry Lewis. Martin himself alluded to this in the March 7, 1964, episode, thanking Jerry for "building me this theater". He enjoyed the gig enough to subsequently agree to star in his own variety series, The Dean Martin Show, which premiered on NBC in the fall of 1965 and ran for nine years.)
Les Brown and his Orchestra served as house band for the first season, with Mitchell Ayres and his Orchestra taking over for the remainder of the run. The off-screen announcer for each program was Dick Tufeld. Grey Lockwood served as director for the show's entire run.
The opening set framing the host established a unique show opening with Jim Trittipo's stage set. After the opening, the set transformed into a second set, with set pieces either splitting apart or turning around, and additional flying set pieces dropping in or flying out on camera, as well as scenic theatrical magic act transforming before the camera while each new act was introduced. This novelty was established as The Hollywood Palace's specialty. This opening transition broke the normal scheduled commercial time-slot breaks, with the commercial break occurring far into the show's first 15-minute segment.
A number of popular music performers got their start on the show; among them were The Rolling Stones, who made their first US television appearance on the episode aired June 6, 1964, and The Jackson 5 made their first national television appearance on the October 18, 1969 episode. The folk-rock group We Five performed their hit "You Were on My Mind" within a few weeks of its release in 1965. During their 1964 appearance, the Rolling Stones were repeatedly ridiculed, before and after their performance of Muddy Waters' "I Just Want To Make Love To You", by host Dean Martin. At the time, they were relatively unknown in the United States and a second song recorded in the same session, Buddy Holly's "Not Fade Away", was not shown until the second episode of the second season, hosted by Ed Wynn, that aired on September 26, 1964. The February 25, 1967, edition featured the American television debut of the Beatles' music videos for "Penny Lane" and "Strawberry Fields Forever", introduced by guest host Van Johnson.
The show, as well as all the ABC's Talmadge Main Lot programming, was televised in black and white until September 1965, when color telecasts were begun. The facility was the first color studio renovated by ABC Television on the West Coast, converted during The Hollywood Palace's summer hiatus. Sharing the studio, scheduling Sunday through Wednesday, The Lawrence Welk Show was moved to Vine Street to broadcast in color at the band leader's request, but the Welk Orchestra's size of players-members were forced to be reduced to fit on the stage. Given the orchestra plan, Welk drew a pencil line on the right side of the plan, announcing, "lose them!".