Hello everyone,
I'm having some conceptual misunderstandings regarding non-reducing SDS-PAGE. In this situation, we omit reducing agents such DTT or BME from the loading buffer to preserve disulphide bonds in the proteins' structure. However, in every protocol i've seen, SDS is present and sample heating is still performed. Wouldn't this result in disrupting the disulphide bridges, since we are still denaturing the samples? I know that disulphide bonds are more heat resistant than hydrogen bonds (since they are covalent bonds) and that heating in the presence of reducing agents is only done to facilitate the disruption of those bonds. But I couldn't understand if high temperature alone is sufficient or not to break these linkages.
Thank you kindly for your attention.
Best regards,
Miguel
Disulfide bonds should be stable to heating in the absence of reducing agent.
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