Dear all,I am a PhD student and I want your help about refolding an enzyme after purification.I induced the enzyme by IPTG but it was found to be insoluble so in order to upload it on a nickel column (as it has His-Tag) I use 6M of urea.After purification, I tried to refold it and remove the urea by dialysis it against Tris buffer 50uM but it precipitated directly. I tried to dialysis it against a gradient of urea with Tris buffer and it didn't work.I tried also to refold it by dilution and it precipitated again.can anyone help me to solve this problem.Thank youSemaz
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