My thesis about the production of mouse anti-human FZD-7 monoclonal antibody (against the 33-185 AA of FZD7) by the hybridoma method. I have a problem with mouse immunization by FZD7. In ELISA the maximum optical density for 1:100 dilution is one. I hear it is so much low for the hybridoma process. Human FZD7 has nearly complete homology with mouse except for two amino acids. I see companies produce mouse anti human anti-FZD7 mAb. Some of them use RAT as a host, but human FZD7 has homology in rat too. I don't know in this situation how can I increase mouse immunization?
hi
1- you can conjugated your recombinant protein with KLH
2- you can immunized rabbit parallel to mouse immunization protocol ( production ploy clonal as positive control for thesis )
3- you can design antigenic peptide for 170-220 aa position in FDZ7 ( region with low homo logy between mouse & human )
good luck
what will be your purpose for the antibody?
If the antibody needs to be used in western blot, you could denature the human protein and immunized the mouse, to produce the antibody recognizes the linear epitope.