Chromokinesins are chromosome-bound proteins during mitosis that play multiple important roles in chromosome segregation. The chromokinesin Kid has been shown to be involved in chromosome congression during mitosis and meiosis. Here we have generated a monoclonal antibody specific for the human chromokinesin hKid by immunizing BALB/c mice with a recombinant protein fragment corresponding to the C-terminal 250-amino acid residues of hKid. All five immunized mice responded excellently and gave high, nearly monospecific, antibody titers. One hybridoma, 8C12, was generated from spleen cells of a selected mouse, which recognized hKid on immunoblots and in immunofluorescence experiments. As hKid is an important regulator of chromosome segregation, this monoclonal antibody will be a useful tool for further analysis of this chromokinesin.