Four major non-collagenous bone proteins were localised by single and double immuno- histochemistry during de novo mineralisation and bone resorption. Both osteopontin and bone sialoprotein were localised ahead of the mineralisation front, suggesting that both proteins are necessary for the initiation of bone mineralisation. This supports previous suggestions that bone sialoprotein acts as a crystal nucleator. The role of osteopontin is less certain, but might be related to ensuring that only the right type of crystal is formed. Osteocalcin and osteonectin were not present in areas of first crystal formation, but were present in the fully mineralised matrix. Their role may be to control the size and speed of crystal formation. Osteopontin, bone sialoproteins and osteocalcin (but not osteonectin) were also present at reversal lines. Interpreting this localisation together with information from the literature, the following functions are suggested during resorption: Osteocalcin may act as a chemoattractant for osteoclasts, while both osteopontin and bone sialoprotein may facilitate the binding of osteoclasts via the arg-gly-asp motif.