Protein solublisation

Biology biolo at iss.com
Sun Jan 31 09:58:13 EST 1999


Hi
Use Phospahte buffer 0.2 M witha variety of different pH like 5-7.5

mahmood
mpiraee at is2.dal.ca


Peter Ashton wrote:

> Hello,
>
> I have a mixture of glycoproteins that I have ammonium sulphate precipitated
> from culture supernatants. I have dialysed the samples extensively against
> PBS to get rid of the ammonium sulphate, but I am having extreme problems
> getting them back into solution. I have tried all of the methods that I can
> find including:
>
> 8M Urea (plus sonication)
> 6M Guanidium-HCl (plus sonication)
> SDS-PAGE sample buffer (and boiling for 30 minutes)
> Glacial acetic acid
>
> In every case, the pellet sits there happily at the bottom of the tube and
> completely refuses to go back into solution.
>
> Does anyone have any other methods that I could try?
>
> Thanks
>
> Pete Ashton
> University of York, UK
> pda2 at york.ac.uk






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