After immunopreciptiting a protein with rabbit polyclonal antibody, I check the binded protein with WB. But the primary antibody I used is also from rabbit. The unspecific signal is terrible. Is that because the antibodies I used are raised from same specis? How can I deal this? Thanks very much
Submit your paper to J Biol Methods today!
1 reply to this topic
Posted 04 August 2002 - 04:12 AM
If you rabbit Ig are a problem you can reduce the amount of antibody that you use (usually 1 microgramme is enought) or you can cross-link the antibody on beads of protein A or G, or if it is available buy an antibody already cross-linked to the beads. If you need a protocol to cross-link your antibody email me.