Another IMAC question

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Sat Aug 22 04:31:22 EST 1998


James Fethiere <james at mpimf-heidelberg.mpg.de> wrote:
> I am using a 1.6ml POROS Ni column to purify a protein eluted from a
> FLAG-M1 column with 2mM EDTA.  After a certain time I start to loose
> binding to the Ni column.

IMO this is not surprising because the EDTA will complex the Ni and
remove it from the column. Either look for another chelator, include
a dialysis step or reverse the order of the columns (i.e. Ni first,
M1 then).

(BTW, I'd be curious how much M1 antibody you need for a column. Must
be pretty expensive...)

--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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