On Wed, 22 Dec 2004 13:32:08 -0600, "Andor J Kiss" <akiss at life.uiuc.edu> wrote:
>Hello,
>> I have a problem with some DEAE Sepharose FF that has clumped. I've
>tried regenerating it, tried high salt (2M NaCl) tried neutral buffer,
>neutral buffer + high salt. Nothing seems to work. Amersham tech support
>is terrible.
>>Any suggestions?
Yes. You are dealing with the [mostly denatured] proteins stuck
hydrophobically to the matrix. High salt is of no use here. Try 6 M GuHCl or
0.5-1.0 M NaOH. No harm even in using 0.5 M NaOH + 0.5% SDS. The only
trick is to wash the sepharose extensively after that to get back to the ~
neutral pH and wash out traces of SDS.
DK
