Purification of proteins in denaturants
mbausher at MAGICNET.NET
Tue Sep 17 14:27:13 EST 1996
Your discription of the problem was brief, but you can use free
solution IEF at very high Urea concentrations (8 M for the Rotofor),
but you have to increase the running temp to 15 C. You also should
deionize the Urea . The highest concentration I have used is 4 M.
You also have to deal with the ampholytes later.
Hope this helps,
mbausher at magicnet.net
From: zaphod at mcimail.com
Date: 9/16/96 5:05:22PM
Subject: Purification of proteins in denaturants
Does any one have any good suggestions for purifying proteins in
denaturants (e.g.,6M urea, 4M guanidine)? I have a his-tagged
which I've been able purify to a great extent by Ni affinity,
but I'd like
to have another chromatographic step. The protein is only
soluble in 6M
urea or 4M guanidine.
Sizing columns work poorly in these buffers and we have not
ion-exhange resin to which the protein will bind in these
Blake Middleton Internet: zaphod at mcimail.com
MCI ID # 465-2423 Bitnet:
zaphod%mcimail.com at cunyvm.cuny.edu
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wasn't previously aware of." --Arthur Dent
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