Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Most SDS-samples were reduced using Dithiothreitol or Mercaptoethanol.
When the samples have cooled down after heating 3 min 96°C, normally an alkylation has to be performed.
This alkylation prohibits re-oxydizing of the Disulfid-bridges.
Reoxydizing can end up in big-sized giga-molweight proteins sticking at the anodal slot-wall.
Mostly happening when the sample-volume is very small in its Eppi-cup.

Remedy:
Use the alkylation reaction after the reducing of the SDS-samples:

This reaction is described in EDC’s manuals:
ElphoGels SDS Kit, SDS DryGel Kit

EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)