SASSKIDM@cms.am.cc.reading.ac.uk ("SASSKIDM") (04/14/89)
Steve Hyde should not despair of renaturing his protein from 8M urea. Our experience, admittedly without the 65o heat treatment, is that both antigenicity and enzymic activity can be regained by stepwise reduction of urea concentration in air at 4o. Briefly, we use serial dilution to lower the urea concentration to 2M in two steps and then dialysis to finally eliminate the urea. The buffer contains 1mM EDTA, 0.5% 2-mercaptoethanol and 20% glycerol. Please contact us for more details. Good luck! Mark Gilbert/Chris Skidmore Department of Biochemistry & Physiology University of Reading