| 摘要 |
PURPOSE:To obtain the subject antibody for measuring a polyubiquitin, etc., derived from an antibody-forming cell by immunization of an animal against a polyubiquitinized protein formed in vitro, being specifically reacted with polyubiquitin but not with monoubiquitin. CONSTITUTION:Ubiquitin is reacted with a denatured lysozyme in 100mM tris- hydrochloric buffer solution (Ph 9.0) at 37 deg.C for 1 hour to give a polyubiquitinized protein such as polyubiquitinized lysozyme. The polyubiquitinized protein is administered to BALB/c mouse and the mouse is immunized. After the final immunization, a splenocyte is collected, subjected to cell fusion with a mouse.myeloma cell to give a hybridoma. Then, a clone capable of producing an anti-polyubiquitin is selected from the hybridoma, the clone is cloned into a monoclone. The clone is cultured in a medium to give the objective anti-polyubiquitin.monoclonal antibody which is capable of specifically reacted with a polyubiquitin, but not with monoubiquitin and measuring polyubiquitin in high sensitivity. |
