Bio-Rad Experion Protein Analysis Kits - Manual (Page 46)

7.1 Electrophoresis and Priming Stations
Error Probable Cause Recommended Action
IV Check Failure
(Experion
™ software)
One or more wells are not filled correctly, and
the electrodes are not immersed Fill all sample wells of the chip with blank or replicate samples
Ensure pipets are properly calibrated
Air bubbles are interfering with the electrical
contact in one or more of the wells Stop the run, remove the chip, and use a clean pipet tip to
remove the sample and apply a new aliquot
When pipetting, insert the tip vertically and to the bottom of
the well. Dispense the liquid slowly. Do not expel air at the end
of the pipetting step. Dispense only to the first stop on the
pipet. Positive (reverse) pipetting may also be used
An electrode is loose or disconnected Perform a conductivity test with the cleaning chip or
conductivity chip from the Experion validation kit (catalog
#700-7051). Add 850 µl water to the chip, place the chip in
the electrophoresis station, and close the lid. In Experion
software, select Tools > Diagnose Instrument and select
Conductivity in the Instrument Diagnostics dialog
If the test passes, focus on the sources of error above. If it
fails, contact technical support with the results
Lid is not closed Close the lid
The chip is not primed properly Repeat the analysis using a new chip
Check Seal
(Priming station) The priming seal is worn, torn, or clogged Replace the priming seal (catalog #700-7031)
The chip is not positioned correctly in the
priming station Align the arrow on the chip with the arrow on the platform
The gel-stain solution (GS) was added to the
wrong well for priming The priming well is highlighted on the chip. See Section 3.6,
Prime the Chip
No chip detected Chip not positioned properly, empty wells,
or incorrect reagents Make sure chip is positioned properly and correct reagents
used
Run aborts before all wells
are analyzed The computer power saver options are
causing the runs to abort (this is only a
problem with Experion software prior to
version 3.0) Upgrade to the newest software version
Confirm that all power saver and/or power options are turned
off before starting the assay. Do not let the computer “sleep”
during a run. If the computer goes to sleep, the run will abort
7.2 Experion Pro260 Analysis
Error Probable Cause Recommended Action
Concentrations are not
reproducible (CV > 20%) Sample preparation procedures are
inconsistent Use the same reagents and follow the same procedures
for each sample and each run. Do not modify the
protocol outlined in Chapter 3 to compensate for protein
concentration or sample variability
Peaks were not detected in
the Pro260 ladder An air bubble is at the bottom of the ladder
well or microchannels due to poor priming or
sample loading; air bubbles can interfere with
the contact between the electrode and the
liquid Check the chip for air bubbles after priming and sample
loading:
n After priming: turn the chip over and make sure the
microchannels are filled with gel-stain solution (GS). If
the chip is primed correctly, the wells should be hard to
see. Compare the primed chip to an unprimed chip if
necessary. Do not touch the glass
n After sample loading: tilt the chip and view it from different
angles against a light background
42 Technical Support: 1-800-4BIORAD • 1-800-424-6723 • www.bio-rad.com
Experion Automated Electrophoresis System