raising ANTIBODIES with fusion proteins??

Riccardo Fodde fodde at ruly46
Tue Sep 6 17:49:12 EST 1994


We have made several his-tagged fusionproteins which we used to immunize 
rabbits. The sera (and also affinity-purified antibody) of these rabbits 
recognize on western-blots besides the expected band, also extra bands. Does 
anyone know a method to get rid of these antibodies recognizing the wrong 
bands? We are thinking of making subfragments of the original fusionprotein 
and then affinity purify the sera, hoping the aspecific antibodies don't 
recognize the subfragment but there must be more clever ways to do this?

Besides this request for help I wonder what the current explanation is for the 
generation of these aspecific antibodies. All reactions and suggestions are 
welcome.

Ron Smits
Leiden University



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