histag elution alternative

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Wed Nov 1 14:19:37 EST 2000


"Richard P. Grant" <rpg14 at yahoo.co.uk.invalid> wrote:
:In article <l2bct8.h9j.ln at wpxx02.toxi.uni-wuerzburg.de>, Cornelius 
:Krasel <krasel at wpxx02.toxi.uni-wuerzburg.de> wrote:
:
:> A resin treated with DTT is almost impossible to recharge
:> (I know since I tried :-).
:
:This deserves repeating over in methds-reagents . . .
:

Interesting... While I do not dispute the finding (never tried 
myself), I am puzzled. Why would it be so??? There simply 
isn't anything in NTA-agarose that can be reduced by DTT.
NTA is coupled to agarose through spacer via AFAIR epoxy
chemistry (not affected by DTT), NTA itself is basicaly three 
acetates linked together.  Sepharose is just a crosslinked 
agarose, so it is not affected by DTT either. 

Anyone wants to bet? I say that reduced Ni-NTA agarose 
can be stripped and recharged w/o a problem. 

        - Dima






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