Table 4 General comparison of IPG- and NEPHGE- based 2DE methods
Method | IPG | NEPHGE |
|---|---|---|
Characteristics | ||
Preparation of 1st dimension gels | Commercial gels; easy to prepare for IEF | Home-made gels; preparation requires skills |
Procedure | Simple, easy to use | Complex, requires skills |
Time for analysis | Fast, 2 days | Time-consuming, 5–6 days |
Price | Cheap (Invitrogen) | Moderate (WITAvision) |
Handling of 1st dimension gels | Handling of IPG strips is safe and easy | Gels are fragile, handling requires serious skills |
Reproducibility | Well-reproducible in acidic, poor in a basic zone | Lower in acidic zone, but excellent in basic zone |
Possible problems | Poor reproducibility of basic protein spots, protein capacity is limited | Handling difficulties, missing of some highly acidic protein spots |
Protein capacity, effect of high protein load | Protein capacity is limited, quality of spots is worse at high protein load | Higher protein capacity, than in IPG gels; quality of spots is good at high load |
General characteristic | Simple and easy to use method; ideal for 2-DE of acidic proteins. Drawback is poor reproducibility of basic protein spots. | Method requires skills; excellent for 2-DE of basic proteins. Analysis in acidic zone is satisfactory, but some spots are missed. |