Cooke had 30 U.S. top 40 hits between 1957 and 1964, and a further three after his death. Major hits like "You Send Me", "A Change Is Gonna Come", "Cupid", "Chain Gang", "Wonderful World", and "Twistin' the Night Away" are some of his most popular songs. Cooke was also among the first modern black performers and composers to attend to the business side of his musical career. He founded both a record label and a publishing company as an extension of his careers as a singer and composer. He also took an active part in the American Civil Rights Movement.

On December 11, 1964, Cooke was fatally shot by the manager of the Hacienda Motel in Los Angeles, California, at the age of 33. At the time, the courts ruled that Cooke was drunk and distressed, and that the manager had killed Cooke in what was later ruled a justifiable homicide. Since that time, the circumstances of his death have been widely questioned.

Cooke was born in Clarksdale, Mississippi. He later added an "e" onto the end of his name, though the reason for this is disputed. He was one of eight children of Annie Mae and the Reverend Charles Cook, a Baptist minister. He had a brother, L.C., who some years later would become a member of the doo-wop band Johnny Keyes and the Magnificents. The family moved to Chicago in 1933. Cooke attended Wendell Phillips Academy High School in Chicago, the same school that Nat "King" Cole had attended a few years earlier.

Cooke began his career with his siblings in a group called The Singing Children when he was nine. He first became known as lead singer with the Highway QC's as a teenager joining at the age of 14. In 1950, Cooke replaced gospel tenor R.H. Harris as lead singer of the gospel group The Soul Stirrers. Under Cooke's leadership, the group signed with Specialty Records where their first recording was for the song "Jesus Gave Me Water" in 1951. They also recorded other gospel tracks, such as "Peace in the Valley", "How Far Am I From Canaan?", "Jesus Paid the Debt", and "One More River", among many other gospel songs some of which he wrote himself. Cooke was often credited for bringing the attention of gospel music to a younger crowd of listeners, mainly girls who would rush to the stage when the Soul Stirrers hit the stage just to get a glimpse of Cooke.

His first pop single, "Lovable" (1956) which was a remake of the gospel song "Wonderful", and was released under the alias "Dale Cook" in order not to alienate his gospel fan base; as there was a considerable stigma against gospel singers performing secular music. However, it fooled no one - Cooke's unique and distinctive vocals were easily recognized. Art Rupe, head of Specialty Records, the label of the Soul Stirrers, gave his blessing for Cooke to record secular music under his real name, but he was unhappy about the type of music Cooke and producer Bumps Blackwell were making. Rupe expected Cooke's secular music to be similar to that of another Specialty Records artist, Little Richard. When Rupe walked in on a recording session and heard Cooke covering Gershwin, he was quite upset. After an argument between Rupe and Blackwell, Cooke and Blackwell left the label.

In 1957, Cooke appeared on ABC's The Guy Mitchell Show. That same year, he signed with Keen Records. His first release "You Send Me", (the B-side of a reworking of George Gershwin's "Summertime") spent six weeks at #1 on the Billboard R&B chart. The song also had mainstream success, spending three weeks at #1 on the Billboard pop chart.

In 1961, Cooke started his own record label, SAR Records, with J.W. Alexander and his manager, Roy Crain. The label soon included The Simms Twins, The Valentinos, Bobby Womack, and Johnnie Taylor. Cooke then created a publishing imprint and management firm, then left Keen to sign with RCA Victor. One of his first RCA singles was the hit "Chain Gang". It reached #2 on the Billboard pop chart and was followed by more hits, including "Sad Mood", "Cupid", "Bring it on Home to Me" (with Lou Rawls on backing vocals), "Another Saturday Night" and "Twistin' the Night Away".

Like most R&B artists of his time, Cooke focused on singles; in all he had twenty-nine top-40 hits on the pop charts, and more on the R&B charts. He was a prolific songwriter, and wrote most of the songs he recorded. He also had a hand in overseeing some of the song arrangements. In spite of releasing mostly singles, he released a well received blues-inflected LP in 1963, Night Beat, and his most critically acclaimed studio album Ain't That Good News, which featured five singles, in 1964.

In 1963, Cooke's 18 month old son, Vincent, wandered away from his mother's supervision and drowned in their front yard pool while Sam was away from the home. With their marriage already in trouble largely due to extramarital affairs by both Sam and his wife, Barbara, the distance between them deepened as Sam blamed Barbara for their son's death. Cooke retreated into a deep depression, and asked that no one wear black to the child's funeral. He found his escape in out-of-town performances, which he agreed to at every opportunity.

On the night of December 11, 1964, Sam had withdrawn a considerable amount of cash, reportedly to buy Christmas presents. At Cooke’s request, the manager of the Hacienda Motel in South Central Los Angeles, where he was staying, Bertha Franklin (who had shot and killed a man six months earlier at the same location), made arrangements with a local prostitute named Elisa Boyer, to "scoop" Sam at a local bar. Boyer, 22-years old, was to lure Cooke back to his motel room. As Cooke and Boyer entered the room, Sam was hit from behind and knocked unconscious.

Elisa Boyer retrieved Cooke’s cash and met Bertha in the motel office.

Cooke awoke shortly thereafter, disoriented and without most of his clothes and belongings. He headed towards the motel office and observed the prostitute and motel manager counting his money. Cooke demanded his items returned, and when Boyer and Franklin refused to open the door, Cooke banged until the door fell off its hinges. When Cooke regained himself, the hotel manager, Bertha Franklin, shot him and told Boyer to run and call the police from a phone booth.

At first, Cooke's murder was chalked up as just another unidentified “rapist” killed in Watts. It wasn’t until the following Monday morning that a reporter determined that Sam Cooke was signed into the motel registry as an occupant, in his own name, and that a world-famous singer had been killed under shady circumstances. By that point, the story had not been covered by the media.

Bertha Franklin, manager of the Hacienda Motel, told police that she shot and killed Cooke in self-defense because he had attacked her. Police found Cooke's body in Franklin's apartment-office, clad only in a sports jacket and shoes, but no shirt, pants or underwear. The official police record states that Franklin fatally shot Cooke who had checked in earlier that evening. Franklin claimed that Cooke had broken into the manager's office-apartment in a rage, wearing nothing but a shoe and a sports coat demanding to know the whereabouts of a woman who had accompanied him to the hotel. Franklin said that the woman was not in the office and that she told Cooke this, but the enraged Cooke did not believe her and violently grabbed her, demanding again to know the woman's whereabouts. According to Franklin, she grappled with Cooke, the two of them fell to the floor, and she then got up and ran to retrieve her gun. She said that she then fired at Cooke, in self-defense, because she feared for her life. Cooke was struck once in the torso, and according to Franklin, he exclaimed, "Lady, you shot me", before mounting a last charge at her. She said that she beat him over his head with a broomstick before he finally fell, mortally wounded by the gunshot. Franklin herself also told later that she fired the gun at least 3 times.

According to Franklin and the motel's owner, Evelyn Carr (whose last name is identified by some sources as "Card", rather than "Carr"), they had been on the telephone together at the time of the incident. Thus, Carr claimed to have overheard Cooke's intrusion and the ensuing conflict and gunshots. Carr called the police to request that they go to the motel, informing them that she believed a shooting had occurred.

A coroner's inquest was convened to investigate the incident. The woman who had accompanied Cooke to the motel was identified as Elisa Boyer, who had also called the police that night shortly before Carr. Boyer had called the police from a telephone booth near the motel, telling them she had just escaped being kidnapped.

Boyer told the police that she had first met Cooke earlier that night and had spent the evening in his company. She claimed that after they left a local nightclub together, she had repeatedly requested that he take her home, but he instead took her against her will to the Hacienda Motel. She claimed that once in one of the motel's rooms, Cooke physically forced her onto the bed and that she was certain he was going to rape her. According to Boyer, when Cooke stepped into the bathroom for a moment, she quickly grabbed her clothes and ran from the room. She claimed that in her haste, she had also scooped up most of Cooke's clothing by mistake. She said that she ran first to the manager's office and knocked on the door seeking help. However, she said that the manager took too long in responding, so, fearing Cooke would soon be coming after her, she fled the motel altogether before the manager ever opened the door. She claimed she then put her own clothing back on, hid Cooke's clothing, and went to the telephone booth from which she called police.

Boyer's story is the only account of what happened between the two that night; however, her story has long been called into question. Inconsistencies between her version of events and details reported by other witnesses, as well as circumstantial evidence (e.g., thousands in cash that Cooke was reportedly carrying were never recovered, and Boyer was soon after arrested for prostitution), invited speculation that Boyer may have gone willingly to the motel with Cooke, then slipped out of the room with Cooke's clothing in order to rob him, rather than to escape an attempted rape.

Such questions were ultimately deemed beyond the scope of the inquest, whose purpose was to establish the circumstances of Franklin's role in the shooting, not to determine precisely what had transpired between Cooke and Boyer preceding the event. Boyer's leaving the motel room with almost all of Cooke's clothing, regardless of exactly why she did so, combined with the fact that tests showed Cooke was inebriated at the time, provided what inquest jurors deemed a plausible explanation for Cooke's bizarre behavior and state of dress, as reported by Franklin and Carr. This explanation, in conjunction with the fact that Carr's testimony corroborated Franklin's version of events, and the fact that police officials testified that both Boyer and Franklin had passed lie detector tests, was enough to convince the coroner's jury to accept Franklin's explanation, and return a verdict of justifiable homicide. With that verdict, authorities officially closed the case on Cooke's death.

Some of Cooke's family and supporters, however, have rejected Boyer's version of events, as well as those given by Franklin and Carr. They believe that there was a conspiracy to murder Cooke and that the murder took place in some manner entirely different from the three official accounts.

Cooke's funeral was held in Chicago at A.R Leak Funeral Home, where thousands of fans had lined up for over four city blocks to view his body. In her autobiography, Rage to Survive, singer Etta James claimed that she viewed Cooke's body in the funeral home and that the injuries she observed were well beyond what could be explained by the official account of Franklin alone having fought with Cooke. James described Cooke as having been so badly beaten that his head was nearly separated from his shoulders, his hands were broken and crushed, and his nose mangled.

No concrete evidence supporting a conspiracy theory has been presented to date.

Cooke was interred in the Forest Lawn Memorial Park Cemetery in Glendale, California.

Some posthumous releases followed, many of which became hits, including "A Change Is Gonna Come", an early protest song that is generally regarded as his greatest composition. After Cooke's death, his widow, Barbara, married Bobby Womack. Cooke's daughter, Linda, later married Bobby's brother, Cecil.