Jacob Hamblin was born in Salem, Ohio on April 6, 1819. He was baptized a member of the Church of Jesus Christ of Latter-Day Saints on March 3, 1842, at the age of twenty-two. He married Lucinda Taylor (1823-1858) who was baptized soon afterward. However, when Hamblin proposed moving west, Lucinda refused to go. In February 1849, Hamblin and Lucinda decided to end their marriage, and he continued west without her, taking their four children with him. In September of the same year, Hamblin met and married Rachel Judd, a widow, in Council Bluffs, Iowa. She had two children from her deceased husband, and Hamblin and Rachel had five more children together. In 1857, he married Sarah Priscilla Leavitt (1841-1927); they had nine children. Hamblin also had six children with his last wife, Louisa Bonelli (1843-1931). In addition to the twenty-four children by his four wives, he adopted three Native American children.

Hamblin was called by President Brigham Young to work with the Paiute Indians of Southern Utah. This calling began a lifetime of work with various tribes. He was later called as the president of the Indian Mission.

He made nine missionary visits to the Hopi villages of Northern Arizona and, in the process, reopened the ancient Ute Crossing on the Colorado River. He pioneered the Lee's Ferry Crossing, and in 1862-1863 traveled completely around the Grand Canyon. In 1870, he guided United States government explorer Major John Wesley Powell on a survey of the Grand Canyon. In November of that same year, he was responsible for the negotiation of the Treaty of Fort Defiance, New Mexico.

In 1869, Hamblin moved from Santa Clara to Kanab, Utah, then nine years later he moved to northern Arizona. In 1882, Hamblin moved to Pleasanton, New Mexico where he died four years later in 1886.