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Does a protein sample that has been denatured at 95°C need to be denatured again after being stored at -20?
At the time of protein extraction, I have added DDT to the samples and then boiled it at 95 °C for 10 min to denature the protein, then I store the samples in -20, I wonder if I need to renature the samples before load them when I perform western blot, and what is the effect if the samp
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Jason Valle Answered Sep 28, 2022
university of Nottingham
Denatured protein samples can be stored in -20 for a short period of time, within a few weeks they can not be boiled twice for Western blot, and the results have little impact. If you are not very reassured, you can heat them for 5 minutes before loading, but be careful not to freeze-thaw the samples repeatedly.
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