blue dye column

Warren Gallin wgallin at
Wed Jan 15 12:51:22 EST 1997

In Article <5bj2a0$3l5u at>, klenchin at
(Dima Klenchin) wrote:
>In article <01bc02b5$0d4fc900$0815f88f at>, "Jang Young Lee"
<jylee at> wrote:
>#Currently, I am working with a cibacron blue dye columne to purify a
>#dehydrogenase of my interest. The enzyme does not show activity without
>#NAD. As far as I know, cibacron blue has a relatively strong affinity
>#towards NAD requiring enzyme. To the contrary, the activity was found in
>#non-bound fractoin in my case. Can anyone give me a reasonable explanation
>#on this.
>The reasonable explanation is that dye chromatography remains totally 
>empirical and all the general outlines you can find in books (Blue -> NAD, 
>Red -> ATP, etc) reflect only a _tendency_ for proteins that do bind. 
>Play with Mg2+ concentration ot try other dyes (Red and Green are next to 
>Blue with respect to the enormous number of very different proteins they 

You don't say what you have in your loading buffer.  If you have NAD
present, that could be your problem, since the dye is supposed to bind to
the NAD-binding site.  Just a thought.
Warren Gallin
Department of Biological Sciences
University of Alberta
Edmonton,  Alberta     T6G 2E9
wgallin at

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