I have purified antibody from hybridoma cell culture (grown with FBS in the media). When using these antibodies to coat plates, I can probe the plates with anti-mouse IgG and detect the coated antibody. However, if I run these same purified antibodies on western blot and probe with the anti-mouse IgG, I don't get any signal seen.
I see a heavy and light chain on SDS page but when I run a western blot the bands react with bovine IgG or IgM but not with mouse IgG
This same antibody was able to be used successfully in immunohistochem to light up the antigen in the tissue section.