p50csk is a protein tyrosine kinase (PTK) that has been reported to regulate the activity of other PTKs belonging to the src gene family. Several hybridoma clones that produce monoclonal antibodies (MAbs) directed against recombinant p50csk were established. Five of the clones were analyzed for their ability to recognize native and denatured p50csk protein after undergoing native and denaturing polyacrylamide gel electrophoresis followed by western blotting. In addition, the clones were tested for their ability to immunoprecipitate p50csk and yet maintain tyrosine kinase activity of antibody-bound p50csk. None of the clones cross-reacted with pp60c-src, a PTK that shares with p50csk the homologous SH1 catalytic domain and SH2 and SH3 regulatory domains. These MAbs can be used to study p50csk directly, and its role in regulating members of the src family.