polyclonal antibody purification

Martin Offterdinger a8803349 at unet.univie.ac.at
Tue Jan 28 02:28:44 EST 1997


On 26 Jan 1997 21:09:24 GMT, ageorge at casbah.acns.nwu.edu (ANNE GEORGE)
wrote:

>I have been having problems trying to purify a polyclonal antibody raised in 
>rabbits. The antigen was an e.coli expressed protein. There is a lot of background.
>does anyone out there know what is the best method.
>Thanks
>-- 
>ANNE GEORGE
>Northwestern University, Evanston, IL.   USA
>ageorge at casbah.acns.nwu.edu
>
Dear Anne
I would recommend affinity purification:
Immobilise the purified protein on a column e.g.: HiTrap-NHS activated
from Pharamacia, centrifuge the serum , dilute it 1:20 with PBS apply
it onto the column and do an acidic elution with Glycine/HCl pH=2.7-
afterwards the eluate has to be neutralised.
Good luck Martin



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