Maintenance and Troubleshooting 101
Troubleshooting the Protein Application
7
Symptoms (Protein)
Missing Upper Marker
Back to “Symptoms (Protein)” on page 89
Most probable causes Solution
Upper marker was called incorrectly. Check upper marker assignment. Follow instructions for “Manual Marker
Assignment” on page 96.
Improper preparation of sample buffer
or denaturing solution.
Refer to the Kit Guide for instructions on storage and preparation of the sample
buffer or denaturing solution.
Incompatible sample component.
Some components of the buffer, e.g.
CHAPS, TFA, etc. may interfere with
the upper marker and decrease
sensitivity.
See Kit Guide for a list of compatible buffers and buffer compounds. For an
updated list, refer to the web site www.agilent.com/genomics/bioanalyzer.
If necessary, dilute, dialyze or desalt the sample. It is recommended to perform a
serial dilution with water or compatible buffer to determine the optimal dilution.
Diluted samples are too old. Use diluted samples within one day. Store samples at 4°C when not in use for
more than 1 hour.
Probable causes Solution
Digestion of upper marker by
proteases.
Add protease inhibitor cocktails to cell lysate samples.
Improper denaturation of samples. Use fresh sample aliquot. Heat samples with denaturing solution for 5 minutes at
100°C
Use 0.5 mL tubes for denaturing samples.
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