I need help removing IgG from protein homogenates!
bss3pk at surrey.ac.uk
Mon Jan 13 07:06:10 EST 1997
Jil Tardiff <jtardiff at westnet.com> wrote:
>I could really use a hand with a confounding problem.
>What I would *like* to do is figure out a way to *remove* the IgG from
>the initial protein homogenate. The protein is isolated from whole mouse
>hearts in a low-conc KCl-KPO4 buffer. And yes, I try mightily to wash
>the blood out before homogenization.
>Any and all help, suggestions, advice would be greatly appreciated!
Try, immobilised protein A (won't be complete), immobilised protein G
(better, but still won't catch them all), or immobilised anti-mouse
IgG (make sure it covers all isotypes and recognizes heavy chains
Alternatively, 12% PEG 6000 will precipitate most of the IgG, as will
33% sat. ammonium sulphate. These may also precipitate your protein
too though :(
Hope this helps
More information about the Methods