Protein solublisation

Peter Ashton pda2 at york.ac.uk
Thu Jan 28 05:07:45 EST 1999


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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