Gel Electrophoresis: Proteins

a monoclonal antibody directed against Ki67, purchased from Dako, Glostrup, Denmark. The authors did not recommend the use of this antibody on paraffin wax sections. We have also tested the antibody on sections from routine blocks from human gingiva, tonsils and lymph nodes using different time schedules for microwave pretreatment and application of the primary antibody. Two crucial procedures for detecting Ki67 antigen successfully were identified. Firstly, slides should be pretreated in a microwave oven in citrate buffer (pH 6.0) for over 20 minutes (4 x 5 minutes).2 Secondly, Ki67 antibody (diluted 1 in 100-200) should be incubated overnight at 4°C to produce strong staining. Torp et al pretreated sections by microwaving for only 10 minutes (2 x 5 minutes) and incubated the Ki67 antibody for only one hour. Under these conditions, we could not achieve satisfactory staining on sections of the three tissues described above. Bankfalvi et ar have reported that almost identical results were found with the Ki67 and MIB 1 antibodies. In their method, microwave heating was carried out for 35 minutes (7 x 5 minutes) and the Ki67 monoclonal antibody