histag elution alternative

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Mon Nov 6 04:49:38 EST 2000


In bionet.molbio.methds-reagnts Dima Klenchin <klenchin at facstaff.remove_to_reply.wisc.edu> wrote:
> But, enough theories. Here is an experiment. I took an old column that 
> needs to be regenerated anyway, filled it with 200 mM DTT in TBS
> and in 30 min it turned dark brown-orange. Good, here we have the 
> reduction. Now the only thing that remains is to remove this Ni+ and
> recharge with Ni2+. Surprisingly, it turned out to be _very_ difficult:

Thanks for confirming my findings. Now, should we do a joint publication
in BioTechniques? :-)

> 1 M HCl, upon long "soaking" incubation for 1 Hr at room temp (totally 
> OK for crosslinked agarose),  decolorizes the column turning it into 
> normal colorless/white agarose.

I wasn't so sure about the stability of agarose in HCl, that's why I
didn't try this.

--Cornelius.

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */






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