IPG-Dalt of Very Alkaline Proteins
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Compared to classical two-dimensional electrophoresis (2-DE) with carrier ampholytes (1 ,2 ), the advent of 2-DE with immobilized pH gradients (IPG-Dalt) (3 ) has produced significant improvements in 2-D electrophoretic separation with respect to:
| 1. |
Higher resolution by using well-defined narrow-pH gradients (3 ).
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| 2. |
Improved reproducibility, as was demonstrated by interlaboratory comparisons (4 ,5 ).
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| 3. |
Higher loading capacity for micropreparative runs, which has accelerated spot identification by microsequencing and mass spectrometry (6 –9 ).
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| 4. |
The possibility to resolve—under steady-state conditions—alkaline proteins, which are normally lost by the cathodic drift of carrier ampholyte focusing or separated by nonequilibrium pH gradient electrophoresis (NEPHGE) with limited reproducibility (10 ).
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