Holstein is a German and Danish surname, often used with the nobiliary particle "von", meaning "of", and may refer to:
- Anna Morris Holstein (1824-1901), American organizational founder, civil war nurse, author
- Barry Holstein (born 1943), American physicist
- Betty Holstein, Australian tennis player
- Eli Holstein (born 2004), American football player
- Friedrich von Holstein (1837-1909), German statesman
- Helvig of Holstein (1260–1324), a Swedish queen
- Isaac Holstein (born 1987), Filipino-American basketball player
- Mechtild of Holstein (1220 or 1225–1288), a Danish queen consort
- Theodore Holstein (1915-1985), American theoretical physicist
- Franz von Holstein (1821-1878), German composer