In article <01bc02b5$0d4fc900$0815f88f at hskim.kaist.ac.kr>, "Jang Young Lee" <jylee at bioneer.kaist.ac.kr> 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
bind).
- Dima
