Rapid imaging, using a cooled charge-coupled-device, of fluorescent two-dimensional polyacrylamide gels produced by labelling proteins in the first-dimensional isoelectric focusing gel with the fluorophore 2-methoxy-2,4-diphenyl-3(2H)furanone.
A new method for visualising proteins in two-dimensional polyacrylamide gels was developed. Proteins were labelled with the fluorophore 2-methoxy-2,4-diphenyl-3(2H)furanone (MDPF) while present in the first-dimensional gel after isoelectric focusing and subsequently electrophoresed into the second-dimensional gel. High resolution spot patterns were produced and compared with other methods of visualisation. A new rapid imaging system based on a cooled charge-coupled-device was used to view the two-dimensional fluorescent protein spot patterns. The method allows the immediate and rapid imaging of two-dimensional gels at the end of electrophoresis with no further processing.
Jackson
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P
Department of Clinical Biochemistry, School of Clinical Medicine, University of Cambridge, Addenbrooke's Hospital, UK.
Urwin
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VE
Mackay
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CD
D001426:Bacterial Proteins / Q000032:analysis*; D003720:Densitometry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D005663:Furans; D007525:Isoelectric Focusing; D010781:Photography; D013194:Staining and Labeling
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D005663:Furans; C008394:2-methoxy-2,4-diphenyl-3(2H)-furanone