Born and raised in San Pedro, California and growing up air drumming and using pots and pans as makeshift drums, Matt Chamberlain began playing drums at the age of ten. He was influenced by the local San Pedro music scene and by getting in to all ages shows at the Dancing Waters Club (Minutemen, Black Flag, Firehose) and by listening to the drumming of John Bonham, Ringo Starr, Stewart Copeland, Neil Peart, and Terry Bozzio. Chamberlain received his first set of drums at the age of fifteen, and studied with David Garibaldi, Murray Spivack, Chuck Flores, and Gregg Bissonette. Three years later, he was awarded a scholarship to study at North Texas State University where he studied with Henry Okstel. Chamberlain quit college and started playing professionally in the Dallas, Texas Deep Ellum scene in the mid-1980s.
His first notable work was with Ten Hands, a locally popular Denton, Texas band. Ten Hands briefly enjoyed heaps of acclaim and an impressive following in the band's home state. In 1988, Chamberlain left Ten Hands to join Edie Brickell & New Bohemians, with whom he toured and recorded from 1988 until 1991 when the band broke up. He can be heard playing on the band's 1990 album, Ghost of a Dog. In the summer of 1991, he briefly joined the band Pearl Jam and toured with the band before the release of its debut album, Ten. He can be seen in the band's first video, "Alive". Chamberlain said, "I toured in the van with them, played a bunch of clubs—I had a good time, definitely had a good time. They wanted me to join the band, but Edie Brickell & New Bohemians had just broke up, I had just got off the road after touring for four years straight—I could not imagine doing it again." For his replacement, Chamberlain suggested drummer and friend Dave Abbruzzese. Chamberlain left Pearl Jam to join the house band for Saturday Night Live during the 1991–1992 season where he was able to play with and back up artists such as Johnny Winter, Charlie Musselwhite, Jon Faddis, Steve Turre, Leon Pendarvis and work with comedians Steve Martin, Chris Farley, Mike Myers and Dana Carvey on musical skits.
At the end of 1992, he moved to Seattle, Washington to start the band Critters Buggin with Skerik and Brad Houser. The band has since released six albums. In 1993, Chamberlain toured as a member of the funk band Weapon of Choice. In 1996, he received career boosts after performing on albums by The Wallflowers (Bringing Down the Horse) which he won a Grammy for "best recorded performance" and Fiona Apple (Tidal). In 1998, Chamberlain began working with singer-songwriter Tori Amos, with whom he gained recognition. He performed on her 1998 album, From the Choirgirl Hotel, and since then he has contributed to several of Amos' albums and tours. He has worked on combining live and looped drums, often creating loops on the fly and then playing along to them. Amos has introduced Chamberlain as "the human loop" at her concerts.
In 2005, he released a self-titled solo album on Web of Mimicry Records that he describes as "an imaginary soundtrack to an Asian-Western-Sci-Fi-Horror Movie." He is in a group called the Slow Music Project with Bill Rieflin, Peter Buck, Robert Fripp, Hector Zazou, and Fred Chalenor. In August 2007, Chamberlain and musician Bill Frisell released the debut album for the musical project Floratone. In 2010, Drum Magazine named him one of the 15 Greatest Groove Drummers of All Time.
On March 27, 2014, Chamberlain made his debut drumming for Soundgarden, the first of a string of live shows on the Lollapalooza tour in South America. The band's drummer and friend of Chamberlain, Matt Cameron announced that he would not be touring with the band in 2014, due to prior commitments promoting Pearl Jam's album Lightning Bolt.