In January 1952, talent scout Ike Turner tried to get Jacobs to record for Modern Records while in Helena, but Jacobs was on his way to Mississippi. They played together in Clarksdale.
Jacobs had put his career as a bandleader on hold when he joined Waters's band, but he stepped out front again when he recorded as a bandleader for Chess's subsidiary label Checker Records on May 12, 1952. The first completed take of the first song attempted at his debut session became his first number one hit, spending eight weeks at the top of the Billboard R&B chart. The song was "Juke", and it is still the only harmonica instrumental ever to be a number-one hit on the Billboard R&B chart. The original title of the track file was "Your Cat Will Play", but was renamed at Leonard Chess's suggestion. (Three other harmonica instrumentals by Little Walter also reached the Billboard R&B top 10: "Off the Wall" reached number eight, "Roller Coaster" reached number six, and "Sad Hours" reached number two while "Juke" was still on the charts.) "Juke" was the biggest hit to date for any artist on Chess and its affiliated labels and one of the biggest national R&B hits of 1952, securing Walter's position on the Chess artist roster for the next decade.
Jacobs left Waters's band in 1952 and recruited his own backing band, the Aces, a group that was already working steadily in Chicago backing Junior Wells. The Aces—the brothers David and Louis Myers on guitars and Fred Below on drums—were credited as the Jukes on most of the Little Walter records on which they played. By 1955 the members of the Aces had each separately left Little Walter to pursue other opportunities and were initially replaced by the guitarists Robert "Junior" Lockwood and Luther Tucker and drummer Odie Payne. Among others who worked in Little Walter's recording and touring bands in the 1950s were the guitarists Jimmie Lee Robinson and Freddie Robinson, and drummer George Hunter. Little Walter also occasionally included saxophone players in his touring bands during this period, among them the young Albert Ayler, and Ray Charles on one early tour. By the late 1950s, Little Walter no longer employed a regular full-time band, instead hiring various players as needed from the large pool of blues musicians in Chicago.