affinity purification of Ab using insoluble protein

Dr. Duncan Clark news at hgmp.mrc.ac.uk
Thu Mar 22 05:20:00 EST 2001


In article <v04011700b6deb4674fd1@[144.92.64.179]>, the eminent Michael
L. Sullivan at BIOSCI/MRC Human Genome Mapping Project Resource Centre
wrote
>What I'm not sure
>is how to go about making an antigen column.  I was thinking perhaps I
>could solubilize the protein in SDS containing buffer, then dialize most of
>it away (I think probably some SDS would remain stuck to the protein
>despite dialysis).  What I'm not sure of is whether this protein
>preparation would then be suitable for something like affigel 10 or 15.

I don't suppose there are any coupling procedures that work in Urea or
GuHCl? 

Alternatively could you make a fusion protein (more work I know :( )
then bind the fusion part to it's normal affinity column even in
denaturant i.e. His, GST, MBP etc. 

Duncan
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk




More information about the Methods mailing list