Why can't you use Prot. G to bind out the anti-bodies? If you are having trouble with Prot. G. you might want to try and Hydroxylapatite Column, or a DEAE column. Both of these should bind out your Ab and the prot. should flow through.
FreD wrote:
> Hi all!
> i am a phd student (bordeaux, France;) and i have some problems with
> an IP. My protein is about 30kDa, and the dimer is about 60kDa. So i
> have some problems to see them on SDS-PAGE (thanks light and heavy IgG
> chains!).
> I can't link my antibodies to proteinG-Sepahrose. I heard a system
> with a filter which can absorb the antibodies: bacteriae produce your
> antigen (Myc in my case), you can fix the extract on a filter and
> incubate it with your eluate from the IP. the antibodies will absorb
> themselves on the filter, and with several filters you can eliminate a
> large amount of antibodies...
> Do you know something about this protocol?
> Many thanks for your advice!
> FreD.
> (sorry for my english, i hope you understand me;)
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