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of large protein subunits. The two gel solutions are poured one
upon the other, and glycerol is added to the 16% gel solution
to avoid mixing with the 10% gel solution—for details see
Schägger and von Jagow ( 11 ).
10. The transfer of the BN gel strip onto a second SDS gel dimen-
sion is recommended to be carried out by embedding the gel
strip while casting a sample gel onto the second dimension gel.
This procedure ensures optimal physical contact between the
BN gel lane and the SDS gel of the second dimension. However,
BN gel strips can also be fi xed onto a prepoured SDS gel using
agarose as usually carried out for 2D IEF/SDS PAGE. Physical
contact of the gels might not be optimal, but the time period
between the end of the fi rst and the start of the second electro-
phoresis is shortened.
Acknowledgments
The authors would like to thank Christina Rode, Institute for Plant
Genetics, Leibniz Universität Hannover, for critical reading of the
manuscript.
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