Dendritic cells (DCs) play an important role in vertebrate immunity, but little is known of the molecular events associated with their development from bone marrow (BM). This report describes induction of a signature protein marking BM commitment to DCs. Using a standard procedure, DCs were generated from BM by cultivation in vitro. Appropriate phenotypic monitoring was done primarily by immunofluorescence, and polyclonal antibody reagents were developed against immature DC lysates. Using one specific antibody reagent, we identified, purified, and sequenced a unique cytosolic phosphoprotein DP58 that occurs within 30 min during BM commitment to DCs. Its sequence matches with a computationally predicted Riken cDNA (GenBank Accession No. XP_138799), and a specific anti-DP58 peptide antibody was developed for further characterization. The study suggests that DP58 induction signals distinct pathway(s) leading to early DC progenitors that may be generated and propagated for a short period in vitro.