Immunoprecipitation...

Allison "allison.haggartyREMOVE THIS" at mcgill.ca
Wed Oct 23 09:00:41 EST 2002



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!).

What if you run a gel without reducing agent.  Then the intact antibody
would run at 150K leaving your 30K and 60K bands on their own.

Allison



>
> 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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