Dear Meg,
There is a database of refolding conditions from which you could get some
inspiration.
http://refold.med.monash.edu.au/search.php
The Urea conditions I have seen there mostly use 37C. for
solubilization/refolding and nearly all contain a reducing agent. Have a
look. There are also a great number of other additives people use to
improve things.
Joerg
> Hi all,
>
> We use 8M urea solubilization buffer for our protein in inclusion bodies
> and
> recommended temperature is 10-15º C. but in 8M conc the urea does not
> dissolve and is in crystalline form only, will it have any effect on
> solubilzation efficiency. Our solubilization time is 1 Hr and after that
> we
> centrifuge and use the supernatant for refolding via dialysis. however the
> pellet after centrifugation of solubilzation show presence of our protein
> on
> sds page analysis. what should we do so that the process of solubilization
> is complete and our protein is not lost in pellet.
>
> thanks in anticipation.
>
> meg
>
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