making a monoclonal from scratch... - (Oct/16/2006 )
I need to make a mouse monoclonal to a human transmembrane protein. Not to use for western blotting etc, but for use as a blocking ab in cell culture experiments to assess the function of the protein. So the ab has to be raised against an epitope that is expressed on one of the extracellular loops.
I've already got the full length protein in a vector for overexpression, but I'm thinking that the best way to start would be simply to order some short peptides based on the known extracellular domain sequences & immnize a mouse with that, and see what we get.
Anyone got any ideas or experience with this sort of thing ???
overexpression of your protein is to prefer in an eucaryotic expression system, isn´t it? I would immunize several mice even with the same immunogenic protein or peptide to screen afterwards for the best antibody, so i would try protein and peptide or even different peptides