92 Maintenance and Troubleshooting
7
Troubleshooting the Protein Application
Symptoms (Protein)
Wrong Sizing Result
Back to “Symptoms (Protein)” on page 89
Most probable causes Solution
Incorrect assignment of ladder peaks. Check assignment of ladder peaks. For details, please refer to the Online Help or
Users Guide.
Incorrect assignment of upper and/or
lower marker.
Store sample buffer and denaturing solution according to the instructions given
in the Kit Guide.
Check assignment of markers. Follow instructions for “Manual Marker
Assignment” on page 96.
Ladder degraded. Use diluted ladder within one day. Store ladder at 4°C when not in use for longer
than 1 hour.
Probable causes Solution
Improper denaturation of ladder. Use fresh ladder aliquot. Heat ladder for 5 minutes at 100°C. Use 0.5 mL tubes for
denaturing.
Least probable causes Solution
Incomplete reduction of samples. Due to disulfide bonds, some proteins will not migrate according to their
molecular weight if they are not reduced properly. Proteins will migrate higher
than the expected molecular weights.
Check preparation of denaturing solution described in the Kit Guide.
Protein characteristics Glycosylation and other post-translational modifications may disturb micelle
formation around the protein. The proteins will migrate higher than the expected
molecular weights. This effect is reproducible.
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