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2-D Electrophoresis and 2-D DIGE

Life Sciences Forums

Two-dimensional electrophoresis (2-D electrophoresis) was first introduced by O'Farrell and Klose in 1975 and is now a widely used method for the analysis of complex protein mixtures extracted from cells, tissues, or other biological samples.

The technique separates proteins according to two independent properties in two discrete steps: (1) Isoelectric focusing (IEF), which separates proteins according to their isoelectric points (pI), and (2) SDS-polyacrylamide gel electrophoresis (SDS-PAGE), which separates them according to their molecular weights (MW).

The power of 2-D electrophoresis as a biochemical separation technique has been recognized since its introduction. Its application, however, has become increasingly significant as a result of a number of developments in separation techniques, image analysis, and protein characterization.

Applications include:

  • Proteome analysis
  • Cell differentiation studies
  • Detection of disease markers in research
  • Cancer research

 

2-D DIGE

2-D Fluorescence Difference Gel Electrophoresis (2-D DIGE) is a variant of 2-D electrophoresis that addresses some of the drawbacks with the original technique, mainly the variation between gels that often leads to multiple repeat runs. This technique has the advantage of running an internal standard as part of each separation, and also allows you to run up to two test samples simultaneously, saving time and labor as well as providing more accurate and reliable results.

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