Appendix B. Troubleshooting - BIO RAD Trans-Blot Turbo Instruction Manual

Appendix B. Troubleshooting

For more information on troubleshooting, see the Bio-Rad Protein Blotting Guide: A Guide to Protein
Detection and Blotting, bulletin 2895. This can be downloaded from our website, www.bio-rad.com, as
a PDF file, or call Technical Support at 1-800-424-6723.
Symptom/Problem
Poor or no electrophoretic transfer;
excessive protein remains in the gel
(as detected by staining the gel after
transfer)
Poor or no electrophoretic transfer;
little or no protein remains in the gel
(as detected by staining the gel after
transfer)
Blot sandwich exceeds 70
transfer
Cassette does not stay in place in
the instrument bay
POWER FAILURE! PRESS ANY KEY
TO CONTINUE is displayed on the
screen
Trans-Blot ® Turbo™ Blotting System
18
Transfer time may be too short. High molecular weight proteins
(>150 kDa) may require increased transfer time.
Change the percentages of the components for handcast gels. The
Bio-Rad Protein Blotting Guide: A Guide to Protein Detection and
Blotting, bulletin 2895, has extensive information on casting acrylamide
gels.
The wrong protocol relative to the gel combination was used for the
transfer. Refer to Table 1 for acceptable combinations of gels that can be
combined in a single run.
There may be an excessive amount of protein loaded on the gel. Reduce
the amount of protein on the gel.
While the preprogrammed protocols efficiently transter most proteins,
some proteins may require further optimization. Increase the transfer time
in 1 min increments. Change voltage and current if required.
The blotting sandwich may be assembled in the wrong order. See Figure
8 for the correct assembly.
The detection system is not working or is not sensitive enough. Include
proper antigen controls to test detection kit sensitivity or use a total
protein stain or Criterion Stain Free™ system to detect protein transfer to
the membrane prior to immunodetection.
ºC after
An incorrect program for the gel combination was used. Refer to Table 1
for the acceptable combinations of gels that can be combined in a single
run.
Reduce run current, voltage, or time.
Make sure that the dial of the cassette lid is in the Lock position and the
lid pins are engaged. The dial should be facing up when the cassette is
inserted into the bay.
The magnetic interlock may be damaged or have failed. Contact
Technical Support for assistance.
Power to the instrument was interrupted during a run. The run is aborted;
it will need to be restarted.
outage, and restart the instrument.
Suggested Corrections
Shut off the power switch, correct the power