There are not as many signs of early human habitation in Pulloxhill as in some other parishes in the county. This may be because the parish is some distance from a major navigable river.

The Bedfordshire Historic Environment Record [HER] contains information on the county's historic buildings and landscapes and summaries of each entry can now be found online as part of the Heritage Gateway website. The earliest evidence for human activity comes from a pit found just off Flitton Road [HER 17794]. This dates to the late Neolithic or early Bronze Age. A small excavation (13 metres by 3 metres) uncovered a north-south aligned ditch containing late Neolithic to early Bronze Age pottery. A large pit containing post medieval pottery was also excavated and is probably associated with nearby earthworks. Another Bronze Age monument is a ring ditch to the north-east of the village [HER 18030]. This is identifiable as a crop mark. Ring ditches are usually interpreted as the remains of round barrows.

In the 20th century human remains were found in the garden of a house on Barton Road [HER 15854]. The bones have been dated to the Iron Age and were probably disturbed by deep ploughing. Crop marks visible south-west of Sand Lane [HER 16646] show a feature with curved lines, perhaps part of a prehistoric enclosure.

The Viatores are a group dedicated to finding Roman roads in the modern landscape. In 1964 they suggested a route from Luton through Barton-le-Clay, Streatley, Harlington, Pulloxhill, Flitton and Greenfield, Flitwick, Maulden, Ampthill and Millbrook to Marston Moretaine [HER 5020] but there is no definite evidence to suggest there was a Roman road in this area.