Coupling proteins in presence of SDS?

David Nunn David_Nunn at qms1.life.uiuc.edu
Fri Apr 15 14:06:39 EST 1994


	Life, ;^) , would be so much easier if it were possible to couple
proteins to a solid support in the presence of SDS. I am interested 
in this for the purpose of coupling SDS-solubilized proteins, that are
overexpressed as inclusion bodies, to a solid support for use as an
immunoadsorbent.
	Most of the proteins I am working with are normally cytoplasmic membrane
proteins that don't renature without a detergent present. What would be
ideal would be to purify these overexpressed proteins on a SDS prep gel
and then directly couple them to a column, removing the detergent
post-coupling without worrying too much if they precipitate on the column
(flow rate problems aside).
	What I have done so far is blot the proteins to PVDF paper, fragment the
paper
in a Waring blender, and use this material as an immunoadsorbent to purify 
specific antibodies. It works alright but not great.
 Anybody know of a coupling procedure that can be done in the presence of 
SDS or something comparable to PVDF that doesn't care too
much if detergent is around?

David Nunn
Department of Microbiology
University of Illinois
Urbana, IL 61801
(217) 333-6131



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