Step Four
Overlay the gel solution with water or water-saturated iso-butanol. Monitor for leaks at this point and allow the gel to polymerize for at least one hour. If the gel is to be used to isolate a protein for the purpose of chemical sequencing we recommend that the gel be allowed to polymerize overnight to ensure maximum polymerization.
Unpolymerized acrylamide can react with primary amino groups and thiols and artificially block the amino terminus of the protein protein. In addition, free radicals left in the gel can catalyze other side-chain modifications that may be deleterious to proper sequencing.
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