We all know that heat is energy, and if too much energy is applied to anything, it is denatured. pH is a denaturing agent since it changes the protein, essentially by using Hydrogen (part of the bonds) as a counteract. Amphiphilic detergents are not beneficial for proteins because they’re responsible for breaking covalent bonds. Mercaptoethanol, as the name says, has ethanol, which reduces proteins into nonexistence.