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
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