PURPOSE: To obtain a mouse monoclonal antibody reacting specifically with P-tyr in cell protein by using P-tyr conjugated protein for immunization of an animal.
CONSTITUTION: Phosphotyrosine (P-tyr) and synthetic peptide or natural protein is brought into reaction and are conjugated. The resulted P-tyr conjugated synthetic peptide or P-tyr conjugated natural protein is intracutaneously injected into a female mouse to immunize the mouse. The spleen cells are drawn from the immunized mouse and are fused with myeloma cells by using polyethylene glycol. The resulted hybridoma is cloned by limiting dilution method and is propagated in the peritoneal cavity of the mouse and is finally propagated as the ascites. The immunoglobulin is settled from the resulted ascites liquid and the settled immunoglobulin fraction is drawn as the antibody for the P-tyr.
IHARA SEIJI
OGAWA RYOICHI
WATANABE YASUSHI
