Generation and characterization of monoclonal antibodies to the Src-familykinase Hck

Hck, a member of the Src-family of protein tyrosine kinases, is expressed p rimarily in hematopoietic cells of the myeloid and B-lymphocyte lineages. H ybridoma cell lines were established that secrete monoclonal antibodies (MA bs) to Hck. Three of the MAbs were extensively characterized and designated H7, H34, and H42. The MAbs H7 and H34 recognized an epitope within the SH3 domain of Hck, while the epitope recognized by the H42 MAb resides within the Unique domain. All three MAbs specifically recognized the p59 and p56 i soforms of Hck in transiently transfected 293T cells and in a murine macrop hage cell line. Notably, the antibodies did not cross-react with other Src- family kinases tested. Under native conditions, the MAbs H34 and H42 effici ently immunoprecipitated Hck from transfected cells. Both MAbs were also su ccessfully used for the immunofluorescent staining of Hck in intact cells. Thus, the MAbs described herein should be useful in studies of Hck function and expression.