strong blue sepharose (?)
Jang Y Lee
Simcity3 at chollian.dacom.co.kr
Tue Jun 24 19:18:47 EST 1997
Make sure that you measure the protein content by the method other than
absorption at 280 nm if you decide to elute with NAD or ATP becuase they
strongly absorb the light at the same wavelength!
Dr. Duncan Clark wrote:
> In article <33AA441B.678B at comp.kbsi.re.kr>, Soohyun Kim
> <shkim at COMP.KBSI.RE.KR> writes
> >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
> Have you tried eluting with NAD?
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
> Duncan Clark
> DNAmp Ltd.
> TEl/FAX 01252376288
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