Differential proteome analysis using 2D-DIGE.
Classical two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) allows comparison and -quantitation of proteomes by visualization of protein patterns using gel stains and comparative image analysis. The introduction of fluorescent reagents for protein labeling (difference in-gel electrophoresis or DIGE) has brought substantial improvement in this field. It provides multiplexing of up to three samples in one gel, higher sensitivity compared to normal protein staining methods, and a higher linear range for quantitation. This article gives detailed protocols for 2D-DIGE, including both minimal as well as saturation labeling.
Department of Medical Proteomics/Bionalaytics, Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany. caroline.may@rub.de
Caroline
C
May
Frederic
F
Brosseron
Piotr
P
Chartowski
Helmut E
HE
Meyer
Katrin
K
Marcus
D000665:Ampholyte Mixtures / Q000737:chemistry; D002021:Buffers; D002232:Carbocyanines / Q000737:chemistry; D002457:Cell Extracts / Q000737:chemistry / Q000302:isolation & purification; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000592:standards; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007525:Isoelectric Focusing; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D018892:Proton-Motive Force; D012015:Reference Standards; D013194:Staining and Labeling; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods / Q000592:standards
D000665:Ampholyte Mixtures; D002021:Buffers; D002232:Carbocyanines; D002457:Cell Extracts; D005456:Fluorescent Dyes; D020543:Proteome