The American Collections Galleries feature objects owned by the Milwaukee Art Museum, the Layton Art Collection (housed at the Museum since 1957), and the Chipstone Foundation (a Milwaukee based institution dedicated to furthering scholarship in the decorative arts).

The core of the paintings collection was formed by Frederick Layton in the late 19th century and includes such highlights as Eastman Johnson's The Old Stagecoach (1871) and John Kensett's Lakes of Killarney (1857). The collections of decorative arts are noted for breadth (spanning from the 17th century through the 21st century) and depth (with particular strength in the 18th century). Highlights include the Layton Art Collection's 17th-century turned Great Chair from New York, the Chipstone Foundation's 18th-century Philadelphia rococo Card Table, and the Milwaukee Art Museum's mid-19th-century rococo revival Sofa made by John and Henry Belter of New York.

In the Museum galleries, the paintings, furniture, ceramics, glass, silver and other decorative arts are arranged thematically, rather than chronologically, so that objects are paired in creative and unexpected ways.

A. Kimbel & J. Cabus (New York, New York, active 1862-1882) Desk, ca. 1876	Michael Allison American, New York or New Jersey, active 1800-1833, died 1855 Tall Case Clock, 1800-25
Washington Allston (American, 1779-1843) A Scene in an Eating House, 1813	American "Tobacco Spit" Glazed Jug, ca. 1890
Unknown, American , 19th century [Bearded man with sideburns], ca. 1850	Unknown, American , 19th century [Bearded man], 1840s
Unknown, American , 19th century [Bearded man], ca. 1850	Unknown, American , 19th century [Bearded man], 1840s
Unknown, American , 19th century [Bearded man], ca. 1840s	Unknown, American , 19th century [Bearded man], 1850s