strong blue sepharose (?)

j.r.stephen mbi071 at sysc.abdn.ac.uk
Tue Jul 1 13:33:07 EST 1997


Hi Soohyun,

If none of these NAD-elution methods work, your enzyme may consist of multiple
components that are eluting seperately, so that there will never be any activity
in any one fraction. In this case, you need to pool fractions in the hope it
will re-associate, but it is not fun, and I hope you find an easier answer!

John Stephen
Soohyun Kim (shkim at COMP.KBSI.RE.KR) wrote:
: Dear, everyone

:         I am purifying some enzymes requiring NAD with blue sepharose of
: Pharmacia. I used 50 mM phosphate or 20 mM Tris, pH 7.0 as starting
: buffer and above buffer containing 2 M KCL or NaCl as elution buffer. I
: confirmed the enzyme activity before chromatography, but I could not
: find the activity after chromatography. I used three column volumes of
: elution buffer. Where is the enzyme? How can I detach the guys from blue
: sepharose? Please send me your idea. 
: With best regards.


: Soohyun Kim

: Korea Basic Science Institute



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