Anthony was an American born in Adams, Massachusetts, the daughter of Quakers. Susan B. Anthony was the second born of eight children in a strict Quaker family. Her father, Daniel Anthony, was a stern man, a Quaker Abolitionist and a cotton manufacturer. He believed in guiding his children instead of directing them. He did not allow them to experience the childish amusements of toys, games, and music, which were seen as distractions from the “Inner Light”. Instead, he enforced self-discipline, principled convictions, and belief in one's own self-worth. Susan was a precocious child and she learned to read and write at the age of three. In 1826, the Anthonys moved from Massachusetts to Battensville, N.Y. where Susan attended a district school. When the teacher refused to teach Susan long division, Susan was taken out of school and taught in a "home school" set up by her father. A woman teacher named Mary Perkins ran the school. Perkins offered a new image of womanhood to Susan and her sisters.

Anthony was independent and educated and held a position that had traditionally been reserved to young men. Ultimately, Susan was sent to boarding school near Philadelphia. Susan taught at a female academy, called Eunice Kenyon's Quaker boarding school, in upstate New York from 1846-1849. After, she settled in her family home in Rochester, New York. It was here that she began her first public crusade on behalf of temperance. While in Rochester, she attended the Unitarian Church.

Anthony was very self-conscious, both of her looks (one eye always pointed slightly outwards) and of her speaking abilities. She long resisted public speaking for fear her speech would not be good enough. However, throughout her lifetime, Anthony worked endlessly. She traveled thousands of miles each year throughout the United States and Europe giving speeches on suffrage (75 to 100 speeches per year for 45 years). She traveled by carriage, wagon, train, mule, stagecoach, ship, ferry boat and sleigh. Anthony died at Rochester, New York, on March 13, 1906 and is buried there in Mount Hope Cemetery. Anthony is known as "The Mother of us all."

In the decade preceding the outbreak of the American Civil War, Anthony took a prominent part in the anti-slavery and temperance movements in New York, organizing in 1852 the first woman's state temperance society in America. In addition, she attended her first women's rights convention in Syracuse in 1852. In 1856 she became the agent for New York state of the American Anti-Slavery Society.

In 1851, Anthony met Elizabeth Cady Stanton. They were introduced, on a street in Seneca Falls, by mutual acquaintance Amelia Bloomer, also a feminist. The two women were to remain close friends and colleagues for the remainder of their lives, although unlike Anthony, Stanton wanted to push a broader platform of women's rights than suffrage.

After 1854, Anthony devoted herself almost exclusively to the agitation for women's rights, and became recognized as one of the ablest and most zealous advocates of complete legal equality, and as a public speaker and writer. She was also active in zealously opposing abortion, then seen as an imposition of men onto women.

From 1868 to 1870, Anthony was the proprietor of a weekly paper, The Revolution, published in New York City, edited by Stanton, and having as its motto: "The true republic — men, their rights and nothing more; women, their rights and nothing less."

In 1869, Anthony and Stanton founded the National Woman's Suffrage Association, an organization dedicated to gaining women the right to vote. Anthony was vice-president-at-large of the National Woman's Suffrage Association (NWSA) from the date of its organization until 1892, when she became president.

In the early years of the NWSA, Anthony made attempts to unite women in the labor movement with the suffragist cause, but with little success. Along with Stanton, she was a delegate at the 1868 convention of the National Labor Union. Anthony alienated the labor movement not only because suffrage was seen as a concern for middle-class rather than working women, but because she openly encouraged women to achieve economic independence by entering the printing trades, where male workers were on strike. Anthony was part of the National Labor Union for a while but then was expelled over this controversy.

For casting a vote in the presidential election held on November 5, 1872, as she asserted the Fifteenth Amendment to the United States Constitution entitled her to do, Anthony was served a warrant on November 18 and was eventually fined $100 on June 18, 1873. She never paid the fine. She was defended at trial by Matilda Joslyn Gage, who asserted that it was the United States that was truly on trial, not Anthony.

In collaboration with Stanton, Matilda Joslyn Gage, and Ida Husted Harper, Anthony published The History of Woman Suffrage (4 vols., New York, 1884–1887). Anthony was also a friend of Josephine Brawley Hughes, an advocate of women's rights and of alcohol abolition from Arizona.

In 1890, Anthony orchestrated the merger of the NWSA with the American Woman Suffrage Association, creating the National American Woman Suffrage Association. Anthony's strategy for suffrage was to unite the suffrage movement where possible, and to focus on the goal of gaining the vote, leaving aside other women's rights issues. Her pursuit of alliances with conservative suffragists created long lasting tension between herself and more radical suffragists such as Stanton. Stanton criticized this stance, writing that Anthony and Lucy Stone, leader in the more conservative American Woman Suffrage Association (AWSA). "They do not see woman's religious and social bondage." Anthony argued to Stanton, "We number over 10,000 women and each one has opinions...we can only hold them together to work for the ballot by letting alone their whims and prejudices on other subjects."

The controversial merger occurred after Anthony created a special National Woman Suffrage Association executive committee to decide whether they should unite with the AWSA (using a committee instead of a full NWSA vote went against the NWSA constitution). Gage (a prominent member who opposed the merger) was denied funds to enable her to attend the NWSA convention leading to these decisions, motions to make it possible for members to vote by mail were strenuously opposed by Anthony and her adherents, and the committee was stacked with members who favoured the merger (two who decided against it were asked to resign).

The union of the two organizations effectively marginalized radical elements of the movement, including Stanton. Anthony pushed for Stanton to be voted in as the first NAWSA president, and stood by her as Stanton was belittled by the large conservative factions within the new organization.

Susan B. Anthony was honored as the first real (non-allegorical) American woman on circulating U.S. coinage with her appearance on the Anthony dollar. The dollar coin, approximately the size of a U.S. quarter, was minted for only four years, 1979, 1980, 1981, and 1999. Anthony dollars were produced at the Philadelphia and Denver mints for all four of these years, and at the San Francisco mint for all production years except 1999.