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 10-15ยบ 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 >
