How can I elute antigen from proteinG+antibody+antigen complexes? - (Nov/29/2006 )
I want to just elute proteins from protein-antibody-proteinG beads without any antibody. What kind of buffer should I use to elute protein and keep antibody still attaching on protein G beads?
Normally, when you elute the protein from the protein G beads, you will also elute the antibody.
Therefore you will need to couple the antibody to the beads using kits, so that only the protein will be eluted.
Hope this help.
I agree with Minnie Mouse. However coupling can cause a whole new slew of problems such as masking the antigenic site that your protein binds to. Also, coupling usually isn't 100% so you may still get some of the antibody eluting. It's still worth a try but just be aware of the problems associated with it.
what is it for? maybe there are some other ways to avoid problems with the eluted antibodies.
are you purifying antigens, or do you want to do immunoprecipitation for western-blot?