Generation and application of a monoclonal antibody that detects a wide variety of protein tyrosine kinases.
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in the human genome, as evidenced by its reactivity with the recombinant Src tyrosine kinases whose subdomain VIB had been replaced by those of the other PTKs.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.
Sugiyama
Yasunori
Y
Department of Life Sciences, Faculty of Agriculture, Kagawa University, Japan.
Sueyoshi
Noriyuki
N
Shigeri
Yasushi
Y
Tatsu
Yoshiro
Y
Yumoto
Noboru
N
Ishida
Atsuhiko
A
Taniguchi
Takanobu
T
Kameshita
Isamu
I
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
D000911:Antibodies, Monoclonal; D010455:Peptides; D011505:Protein-Tyrosine Kinases