His-tagged proteins

Andrew Leech AndrewLeech at see.sig.for.email
Fri Mar 6 13:45:20 EST 1998

In article <mbzrl-060398115348 at pmbfjd0.nottingham.ac.uk>, 
mbzrl at mbn1.biochem.nottingham.ac.uk says...

>Has anyone experienced his-tagged proteins apparently 'disappearing' on
>FPLC gel filtration columns (eg Superose 6 or 12 columns). I assume they're
>coming out of solution, but it's happened now for the last 2 his-tagged
>proteins I've tried to purify but doesn't happen for the GST-tagged
>equivalents.... Very strange!
>Thanks in advance

If you are running your gel filtrations at low ionic strength, the protein
may be sticking to residual charged groups on the column matrix. Try adding
100mM salt to your buffer.


Email: a dotte leech atte uea dotte ac dotte uk - U-no-Y!
Andrew Leech                 *All opinions personal
Biological Sciences          *
University of East Anglia    *Economics is the eugenics of
Norwich, UK                  *the late 20th century.

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