In order to produce markers of melanoblasts and melanocytes, we immunized a mouse with trunk neural crest cells differentiated into melanocytes in culture. We obtained three monoclonal antibodies which react both with pigmented and unpigmented cells in epidermis and in neural crest cultures. These markers are characteristic of distinct differentiation steps. One of them, melanoblast/cyte early marker (MelEM) monoclonal antibody (Mab) detects melanoblasts as soon as they have reached the subectodermal mesenchyme, while the two others, Mel1 and Mel2 Mabs, detect antigens present in melanosomes and promelanosomes later in the differentiation process. Moreover, MelEM is a specific marker of melanoblasts/cytes deriving exclusively from neural crest and not of those from the neural retina. The antigen recognized by MelEM Mab is a protein of Mr 26,000. The epitopes recognized by Mel1 and Mel2 Mabs are carbohydrate moieties carried by glycoproteins of Mr 123,000 and 85,000, respectively. © 1993 Academic Press, Inc.