Jan van der Lee
baslee at kabelfoon.nl
Wed Aug 26 01:50:43 EST 1998
I have had my problems as well using Sepharose columns.
My solution : add some ethanol or glycerol to your buffer system (5-15%
is usually sufficient) to prevent binding to the Sepharose.
For most (not all) of my separations this worked.
Sham Nair wrote:
> Hi Netters, I am trying to purify a calcium-dependent lectin
> from the serum of an invertebrate. After initial affinity
> chromatography (using N-acetyl-D-galactosamine coupled to
> agarose and eluting with EDTA), I tried to separate the
> mixture of proteins in the eluate by size exclusion
> chromatography. The problem was that I lost the lectin during
> this latter process. I presumed that the lectin was binding
> to the chromatography media (Sepharose) and tried to carry out
> the separation with EDTA in my lectin suspension, but this did
> not help. Any suggestions? Thanks in advance.
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