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2D- Electrophoresis

September 21, 2020

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.

[First Dimension]
[Second Dimension]
[2D Troubles]

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:

[Acidic Peptides]
[Basic Peptides]

Appl.Note: One Dimensional and Double Native Electrophoresis of Peptides and Proteins

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

[elpho book]