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 <firstname.lastname@example.org>, the eminent Michael
L. Sullivan at BIOSCI/MRC Human Genome Mapping Project Resource Centre
>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
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.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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