Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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As first dimension an IEF is done. The proteins were separated according to their isoelectric points.
Because a immobilized pH-gradient is more reproducable the IPG-strips are commonly used as first dimension. These strips run denaturated with 8 M Urea so that also the non water-soluble proteins will be displayed.

The proteins within their gel-strips were equilibrated with the normal SDS-sample buffer.
Then as second dimension a SDS-electrophoresis is done. Depending of the amount of different spots the second dimension is done also in large sized gels.

The second dimension functions vertically and als horizontally.

If IEF in IPG-strips is not the optimal method to achieve a good separation, f.e. basic proteins or peptides the two native electrophoresis’ could be done as first dimension:

Pictures from the book “Electrophoresis in Practise” by Dr. R. Westermeier

The 2D-electrophoresis schematically

The 2D-electrophoresis is a very complex field, a overview can be obtained by reading 2 important books of Dr. Reiner Westermeier

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