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Heating to denature and reduce protein: same temperature?

SDS-PAGE reduction denaturation SDS DTT

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#1 Michael Starr

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Posted 18 March 2016 - 12:34 PM

I read the Wikipedia article on PAGE and it says that denaturation can happen at 60oC, whereas reduction requires 100oC. We just do 10 minutes at ~70oC and the reduction seems to work.

 

What is your experience with this? Also, if you heat too long/high, proteins can aggregate. Since it works for me, I am inclined to believe it is working. But maybe it is partial reduction? Is 100oC really required for complete reduction?



#2 mdfenko

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Posted 19 March 2016 - 03:15 PM

maybe the article meant 100C is required for reduction without the addition of reducing agent and ionic detergent.

 

my experience (and much of the literature) suggests that denaturation (and reduction) can be complete within 10-20 minutes at 60-70C, with greatly reduced possibility of aggregation.


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