Establishment of "one-piece" large-gel 2-DE for high-resolution analysis of small amounts of sample using difference gel electrophoresis saturation labelling.
Large-gel two-dimensional gel electrophoresis (2-DE) is the method of choice for high-resolution proteome analysis of complex protein mixtures. Until now, however, the advantages of large 2-DE in combination with multiplexed fluorescence dye protein labelling has been complicated by the separate handling and analysis of the second-dimension gels. Therefore, we adapted the large 2-DE procedure allowing us to run "one-piece" large 2-DE gels (40 cm x 30 cm) in the second dimension for high resolution proteome analysis. Here, we show that in combination with fluorescence dye protein saturation labelling "one-piece" large 2-DE enables analysis of small amounts of sample (3 microg protein) for high-resolution proteome analysis.
Sitek
B
B
Medizinisches Proteom-Center, Ruhr-Universität Bochum, ZKF 1.43, Universitätsstrasse 150, 44801, Bochum, Germany.
Sipos
B
B
Pfeiffer
K
K
Grzendowski
M
M
Poschmann
G
G
Hawranke
E
E
Köper
K
K
Klöppel
G
G
Meyer
H E
HE
Stühler
K
K
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D020543:Proteome / Q000032:analysis* / Q000737:chemistry
D005456:Fluorescent Dyes; D020543:Proteome