A crude purification prior loading the metal-chelating column (like ion
exchange chromatography) can help and speed up the binding to the column
(remark not specific to Fos-cholin). Fos-choline is, contrary to
popular belief, a quite harsh detergent and this may affect the
purification yield and integriry of the protein.
HTH
Daniel
Le 13/07/2010 00:59, Pascal Egea a écrit :
Dear All,
I apologize for the not strictly crystallography-related query.
I am currently purifying several membrane proteins solubilized in
fos-cholines detergents and I consistently observe a significant loss of
protein at the binding step (done in absence of imidazole). Has anyone
else experience the same quite systematic (so far in my hands) problem
with this class of detergents.
I would appreciate any comments or advices from biochemists that face(d)
the same situation.
Thanks in advance
--
Pascal F. Egea, PhD
