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
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