Dye-ligand chromatography....

Debashis from San Diego debmukh at scripps.edu
Fri Sep 28 15:32:04 EST 2001


Hi all,

Hope this newsgroup might help solve our problem.

We have been trying to purify (and subsequently crystallize) a cytosolic
kinase from a hyperthermophile, the gene being cloned and overexpressed in
E.coli. Affi-Gel Blue was found to be most suitable for chromatography and
everything looks great upto the sample loading stage. Then the panic
starts as the protein never comes out even with 3M KCl in the eluting
buffer. It seems something is wrong at this high salt conc. I was thinking
of using NAD+ as elutant.

Any idea, help, suggestion, reference (as I'm a novice in the field) would
be highly appreciated.

Thanks,
Deb 

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                 Debashis Mukhopadhyay, Research Associate
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