I am investigating a protein in human serum that denatures at 4degrees C over time. I now want to try and renature the protein to its native conformation, for this I will try thermal renaturation at ambient temperature and 37degreesC. Does anyone have any experience/suggestions on what else to try?
While you are renaturing protein, it may form aggregates. So to avoid it, add chaotrophic agents at low concentrations and try at different concentrations also
Good luck
The changes in the proteins that you ascribe to denature is more likely a result of proteolysis. Employ proteinase inhibitors.
The protein is stable over the same time period at ambient lab temperature (approx 19-21degC) yet we see a decrease in the assay signal intensity when stored at 4degC. The assay we use to measure is an immunoassay, therefore structural recognition of the protein epitope for signal generation. I suspect that storage at 4degC causes a conformational change in the protein so that the epitope is altered leading to reduced assay result. As it is stable at room temperature, I assume this would rule out proteolysis being the cause.
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