Preparation of antiserum to human C9 with a new immunization method using trace amount of immunogen

Objective To establish a simple and effective immunization protocol to prepare antiserum to human C9 in mice. Methods After characterized purity of C9 with high performance capillary electrophoresis, the immunogen was diluted into appropriate concentration and incubated with NC membrane. Mice were immunized with NC membrane loading C9 protein. The titer of antiserum was detected with undirected enzyme linked immunosorbent assay and the specificity was shown with western blotting. Results The titer of antiserum is about 1×10 -5 in ELISA. Immunological blotting using the antiserum as first antibody showed specific C9 band in sample of normal human serum. Conclusion The immunization method using NC membrane as an immunogen carrier has succeed in getting antiserum with high titer and specificity.