The Chambers Brothers first honed their skills as members of the choir in their Baptist church. This set up ended in 1952 when older brother George was drafted into the army. After his discharge George moved to Los Angeles. The other Chambers brothers soon settled there as well. As a foursome, they began performing gospel and folk throughout the Southern California region in 1954, but they more or less remained unknown until appearing in New York City in 1965.

In the early 1960s, these four brothers from Mississippi, Joe and Willie on guitar, Lester on harmonica, and George on washtub bass, started to venture outside the gospel circuit, playing at coffeehouses that booked folk acts. They played at places like The Ash Grove, one of Los Angeles's most popular folk clubs. It became a favorite haunt of theirs and brought them into contact with Hoyt Axton, Ramblin' Jack Elliott, Reverend Gary Davis, and Barbara Dane. Dane became a great supporter, performing and recording with the brothers. She took them on tour with her and introduced them to Pete Seeger, who helped put the Chambers Brothers on the bill of the 1965 Newport Folk Festival. One of the songs they performed, "I Got It," appeared on the Newport Folk Festival 1965 compilation LP, which was issued on the Vanguard label.

They were becoming more accepted in the folk community, but, like many on the folk circuit, they were looking to electrify their music and become more rock and roll. Guitarist Joe Chambers recalled in a May 1994 Goldmine article that people at the Newport Folk Festival were breaking down fences and rushing to the stage. "Newport had never seen or heard anything like that." After the group finished and the crowd finally settled down, the MC came up and said "Whether you know it or not, that was rock'n'roll." That night they played at a post-concert party for festival performers and went to a recording session of the newly electrified Bob Dylan. Now having gone electric George would trade in his washtub bass for a Danelectro bass guitar.

Shortly after this, the group recorded its debut album People Get Ready.

One night when the group was playing in the Cheetah Club they were met by a man smoking a cigar who said that he was there to bring them to Columbia Records and that's where they met David Rubinson. Actually, Warner Brothers and RCA wanted to sign them as well but wanted to change the bands image and put them in uniforms and have them put down their instruments. This is something that the brothers did not want! In the beginning it didn't seem much better with Columbia.

Columbia president Clive Davis didn't want them to record their song "Time" and said that they didn't record that kind of stuff at Columbia. He also wanted to find a white group to record the song for them. The band said that this would never happen. When they broke the news to David Rubinson he was very heartbroken as he was really looking forward to producing "Time". He found a solution to the problem. He told the brothers that he might lose his job for doing this but they were going to record the song. He told them to come in early for the session. Everything would be ready but they wouldn't have any time to overdub or fix anything on the recording. They went into the LA studio in 1966 and recorded it with Rubinson adding some strange sitar like sounds over the song with some primitive / early Roland Farfisa synthesizers that he found in the studio. It didn't do any good because Columbia refused to put the single out.

Later on when the group had a regional hit with the song "All Strung Out Over You", Columbia allowed Rubinson and the Chambers Brothers to re-record "Time" in 1967. By this time they had expanded "Time" to be the showpiece of their live show. They recorded the new version in one take with all of the effects live in the studio. Rubinson had set up the reverb and reinsertion things with Fred Catero, the studio engineer. Rubinson recalled that it was just one of those incredible magic sessions with all of them reacting to each other and even the trippy time tunnel section in the center of the song was a spontaneous creation.

The band scored its first major hit in 1968 with "Time Has Come Today" (written by Joe & Willie Chambers), from the group's similarly named third album, "The Time Has Come". The song spent five consecutive weeks at #11 on the Billboard Hot 100, just missing the Top Ten.

Later incarnations of the group included session guitarist Steve Hunter (Alice Cooper) and session whiz Stephen Patt (Northwind).[citation needed] , freeing the brothers up to become front men, not just instrumentalists.[citation needed]

However, due to a succession of dishonest promoters and managers, the group found it difficult to build on this success and eventually split up in 1972. Their 1972 album they recorded for Columbia - "Oh! My God" remains unreleased. They reformed in 1974 to record Unbonded and Right Move (1975) for the Avco label. In 1976 they released the final Recorded Live In Concert on Mars for the Roxbury label. They have toured irregularly since.