Cara Baker (crbaker@mail.tcd.ie) wrote: : Has anyone had experience with his-tagged proteins that precipitate : immediately on elution with imidazole? Is there a way to get around : this? I've had the same problem; my protein precipitates when I

[Malini Madiraju]

Cara Baker (crbaker at mail.tcd.ie) wrote:

: Has anyone had experience with his-tagged proteins that precipitate 
: immediately on elution with imidazole? Is there a way to get around 
: this?

Jared (bijgh at zeus.bris.ac.uk ) wrote

I've had the same problem; my protein precipitates when I dialyse away
the
imidazole.  I've found that if I elute the protein by stripping the
column
(with EDTA) I get no precipitation at all.


My question is : When you elute with EDTA, is the eluted portein now
asociated with nickel which may affect its properties and/or activities?

Malini R. Madiraju