Discussion
What is the role of β-mercaptoethanol and bromophenol blue used in separation of proteins by SDS-PAGE ?
Answer: B
β-mercaptoethanol is a reducing agent and bromophenol blue a tracking dye.
In SDS-PAGE, β-mercaptoethanol reduces disulphide bonds so that proteins unfold fully into separate polypeptide chains, which SDS coats with negative charge. Bromophenol blue is a small negatively charged dye that runs ahead of the proteins and shows the progress of the run, so the gel can be stopped before samples run off.
SDS, not mercaptoethanol, is the anionic detergent, and proteins are stained afterwards with Coomassie blue or silver. Mercaptoethanol reduces rather than oxidises. Marker proteins are separate standards of known size, not bromophenol blue.
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