immunoaffinity purification of a protein complex

Haibin Mao hbmao at u.washington.edu
Mon Jan 12 23:02:21 EST 2004


Hello,

We are trying to purify an endogenous protein complex from tissue by 
immunoaffinity column. We want to elute the complex in its native form 
as intact as possible in the absence of the antibody.  So far, we are 
able to purify the full length protein of one subunit of this complex 
in E. coli. We plan to generate polyclonal antibody against this 
subunit and then make an affinity column. We plan to elute the complex 
with the competition method using a lot of the E. coli purified 
subunit. Since we have never done this kind of thing before, we are 
wondering whether this approach sounds reasonable or not to the experts 
out there.

Thanks.


Haibin Mao

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