SDS-PAGE:slower migration after reducing?

logan david.logan at plant-sciences.oxford.ac.uk
Wed Jan 24 08:19:43 EST 1996


John Smith wrote:


>Any ideas why the same sample treated with the same loading buffer 
>(2X conc:
>0.125 M Tris pH 6.8, 4% SDS, 20% glycerol) either (1) without beta-
>mercaptoethanol and without boiling or (2) with BME (5% final conc) 
>and boiling 10 minutes results in a slower migration under 
>reducing/denatureing conditions?

Without reducing the protein you could have S-S bridges resulting in 
a more compact protein thus faster moving protein or you could have 
S-S bridges formed with another polypeptide or heating the sample 
could result in aggregation of polypeptides.




David C. Logan																

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