Henry "Hank" Mobley was born July 7, 1930, in Eastman, Georgia, and raised in Elizabeth, New Jersey, near Newark, in a musical family; he began on piano as a child. At sixteen an illness kept him housebound for several months, and his grandmother bought him a saxophone to occupy the time. Turned away from a Newark music school because he wasn't a resident, he taught himself theory and harmony from books she bought him while teaching himself tenor saxophone at the same time.

He started playing with local bands at nineteen, and within months was working with musicians including Dizzy Gillespie and Max Roach. Roach brought him into the New York jazz scene in 1951, and over the next two years Mobley began composing and recording his own tunes, working through several R&B bands and filling in for two weeks in 1953 with the Duke Ellington Orchestra while saxophonist Jimmy Hamilton recovered from dental surgery. Charlie Parker, on hearing him play, advised him to lean harder into the blues. In April 1953, Roach hired him for The Max Roach Quartet Featuring Hank Mobley (Debut Records DLP-13); Mobley later appeared on two more Roach-led sessions for EmArcy in 1957 and 1958. Around the same period he began working regularly with drummer Art Blakey, and together with pianist Horace Silver, bassist Doug Watkins, and trumpeter Kenny Dorham took part in one of the earliest hard bop recording sessions, released as Horace Silver and the Jazz Messengers (1956, Blue Note). That same Messengers rhythm section backed Mobley on his own 1955 Blue Note debut as a leader, Hank Mobley Quartet.

He recorded steadily through the second half of the 1950s for Blue Note, appearing alongside Lee Morgan, Donald Byrd, Art Farmer, Kenny Dorham, Jackie McLean, Pepper Adams, and pianists Sonny Clark, Bobby Timmons, Herbie Hancock, and Wynton Kelly. He rejoined Blakey's Jazz Messengers in March 1959, recording At the Jazz Corner of the World and a studio session first released decades later as Just Coolin', before leaving that summer; Wayne Shorter took his chair. The 1960s brought his most celebrated work as a leader: Soul Station (1960), generally considered his finest recording, and Roll Call (1960), both with Blakey on drums and the final records the two men made together. The Guardian later gave four of his Blue Note albums from this run, Peckin' Time, Soul Station, Roll Call, and Workout, five stars apiece.

In 1961, while Miles Davis was searching for a permanent replacement for John Coltrane, Mobley spent a brief stint in his band, appearing on Someday My Prince Will Come (on two tracks, playing alongside Coltrane himself), the live In Person: Live at the Blackhawk, and At Carnegie Hall. That year he also cut Workout and Another Workout with a rhythm section of Wynton Kelly, Paul Chambers, and Philly Joe Jones, all Davis band alumni; Another Workout wasn't released until 1985. No Room for Squares (1964) paired him again with Donald Byrd and Lee Morgan, and Dippin' (1965) teamed him with Morgan and pianist Harold Mabern. While serving a prison sentence for narcotics possession in 1964, Mobley wrote the material later recorded as A Slice of the Top, cut in 1966 but not released until 1979. A Caddy for Daddy, Hi Voltage, and The Flip, recorded across the mid-to-late 1960s, drew consistent critical praise, and Reach Out! (1968), featuring guitarist George Benson, was one of his rare accommodations of a more contemporary sound.

A longtime smoker, Mobley was forced into retirement in the mid-1970s by lung problems, and struggled afterward with homelessness and staying in touch with fellow musicians. His last engagements came at the Angry Squire in New York in November 1985 and January 1986, in a quartet with pianist Duke Jordan and guest singer Lodi Carr. He died of pneumonia on May 30, 1986, in Philadelphia, at fifty-five, having also been diagnosed with lung cancer.

Eli Degibri built his entire 2018 album Soul Station as a direct tribute to Mobley's own 1960 record of the same name, a measure of how far Mobley's influence reached into later generations of tenor players who never worked alongside him directly.

Critic Leonard Feather called Mobley the "middleweight champion of the tenor saxophone," a description aimed at a tone that was neither as aggressive as Coltrane's nor as mellow as Lester Young's, and a playing style that stayed laid-back, subtle, and melodic even next to contemporaries like Coltrane and Sonny Rollins. Jazz radio host Bob Perkins later described that sound as "round, throaty, and distinctive." Around the time of his stint with Miles Davis, JazzTimes noted Mobley "retooled his sound" toward a harder edge, a shift some critics trace through his mid-1960s Blue Note run. Critic Stacia Proefrock called him "one of the most underrated musicians of the bop era," and The Guardian's five-star treatment of his four "classic" Blue Note albums praised his "infinite subtlety" and skill as "an ingenious composer." That combination of melodic restraint and compositional craft, played out across dozens of sessions with the era's defining rhythm sections, is what later drew a saxophonist as far removed in time and place as Eli Degibri to build an entire tribute record around Mobley's own Soul Station.