title: Generation and application of a monoclonal antibody that detects a wide variety of protein tyrosine kinases.
abstract: To investigate expression profiles of the entire family of protein tyrosine kinases (PTKs), we attempted to generate an antibody that detects a variety of PTKs. For production of the antibody, antigenic peptides corresponding to amino acid sequences of a highly conserved region (subdomain VIB) of PTKs were synthesized and immunized to BALB/c mice. Among various antigens, a peptide with 11 amino acids, CYVHRDLRAAN, efficiently produced a polyclonal antibody with a broad cross-reactivity to PTKs. We established a hybridoma cell line producing a monoclonal antibody, YK34, which appeared to cross-react with at least 68 PTKs in the human genome, as evidenced by its reactivity with the recombinant Src tyrosine kinases whose subdomain VIB had been replaced by those of the other PTKs. When differentiation of HL-60 cells was induced by 12-O-tetradecanoylphorbol-13-acetate, on Western blotting we observed significant changes in immunoreactive bands with YK34 in HL-60 cell extracts along with changes in the morphology of the cells. These results suggest that the YK34 antibody will be a powerful tool for analysis of a variety of cellular PTKs.
authors.affiliation: Department of Life Sciences, Faculty of Agriculture, Kagawa University, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Sugiyama
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Sueyoshi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shigeri
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Tatsu
authors.forename: Noboru
authors.initials: N
authors.lastname: Yumoto
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Ishida
authors.forename: Takanobu
authors.initials: T
authors.lastname: Taniguchi
authors.forename: Isamu
authors.initials: I
authors.lastname: Kameshita
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D018922:HL-60 Cells; D006801:Humans; D006825:Hybridomas; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D011505:Protein-Tyrosine Kinases / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D010455:Peptides; D011505:Protein-Tyrosine Kinases