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The Catholic community that grew around St Ambrose Parish beginning in the early 1800's was bound by more than a common faith; many members of the parish were also kinfolk. Descended from the Minorcan clan who had lived in St Augustine since the 1700's, these intertwined families migrated inland to find farmland, settling 12 miles southwest of St Augustine at Moccasin Branch.

Priests from St. Augustine served the parish until 1860. Then, Father Henry Peter Clavreul arrived from France to minister to the settlement, and to other Catholic enclaves in southwestern St. Johns County. During this period, the Moccasin Branch faithful attended Mass at a barn on the Solano farm. In 1875, Father Clavreul's successor, Father Stephen Langlade, built the first St Ambrose Church. A skilled carpenter, Father Langlade was architect and contractor on the project. Parishioners provided labor and a quota of five pine trees per family.

From 1881 to 1948, a two-room school operated by the Sisters of St Joesph from St Augustine educated generations of parish children. By the early 1900's, the community had outgrown Father Langlade's hand-hewn building. The "new" St Ambrose Church was finished in 1907. It has served the parish ever since.