Ambler was born on December 24, 1690, in the city of York, England, the seventh of eleven children of John Ambler and Elizabeth Bickerdike Ambler. He was one of only three who lived to maturity. He received little formal education and probably went to work as a young man for his uncle Arthur Bickerdike, a merchant with business connections in London. Bickerdike took Ambler to Virginia in 1716 and established a trading house in Yorktown.
Bickerdike died in 1720, leaving Ambler to operate the business under his own name. Ambler prospered during the 1720s and in 1728 petitioned thefor permission to build a wharf and erect a storehouse on the waterfront. In 1729 he married Elizabeth Jaquelin, the daughter of Edward Jaquelin, a wealthy merchant who owned half of Jamestown Island. At Jaquelin’s death in 1739, Ambler and his wife inherited his property, and between 1744 and 1765 Ambler acquired the remainder of the island through a combination of purchases and lease agreements. As his prosperity mounted, Ambler also purchased land and slaves in Hanover, James City, Louisa, and Warwick counties. Altogether by the time of his death he had accumulated almost 2,000 acres of land and probably 100 or more slaves.
Throughout his life Ambler occupied himself principally as a merchant. He lived in Yorktown and traded in tobacco on his own account or in association with his Yorktown neighbors and fellow merchants, John Norton and his son John Hatley Norton, Philip Lightfoot, and Thomas Nelson and his sons William Nelson and Thomas Nelson. Ambler’s principal London connections were the great mercantile houses of Micajah Perry and Company, Edward and Samuel Athawes, and Edward Hunt and Company. He also traded with Bowden, Farquahar and Kinlock, of London and Edinburgh, and with Farrell and Jones, of Bristol. None of Ambler’s business records survive, but the extent of his trade is evident in the appraisals of his estate following his death. His property, including merchandise on hand and outstanding debts, amounted to about £15,000.
Ambler served as a justice of the peace for York County from November 1724 until November 1737 and as the enumerator of tithables in the county in 1727, 1728, and 1731. The surviving records of Yorkhampton Parish show that he was a vestryman as early as January 1732 and as late as January 1753, and he probably served until his death. He was also collector of the York River customs district for thirty-five years under an appointment made on April 29, 1724. He took the oaths of office in December 1724 and relinquished the office to his son John Ambler in 1759. Ambler had six sons and three daughters, but only three children, all sons, were still alive when he dated his will on August 31, 1765. The date of his death is not known, but family history records suggest that he died about February 1766. His will was proved in York County court on July 21, 1766.