Recombinant protein from cell lysate - (Aug/29/2011 )

Hi, I am trying to purify a recombinant unlabelled glycoprotein protein from a mammalian cell lysate. Can any expert suggest, which are the shortest ways to purify this protein? I am using lectin affinity followed by sephadex 200 for purifying this 130 KDa protein. I tried 2 times i get it on SDS page but no immunoblot activity. I suspect protein gets denatured.
I need some suggestions.

did your unpurified crude extract shows immunoblot activity???

the protein is denatured by sds-page. you need to use an antibody qualified for immunoblotting.

Adrian K on Mon Aug 29 18:34:45 2011 said:

Yes, It blots with monoclonal, even the elute fraction blots but the immunoactivity is much decreased. Also the undesired protein bands sustains in the purified fraction.

Yes, It blots with monoclonal, even the elute fraction blots but the immunoactivity is much decreased. Also the undesired protein bands sustains in the purified fraction.
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