62
Troubleshooting,
Continued
Observation
Cause
Solution
Poor resolution or
smearing of bands
High salt or detergent concentration
in samples
Be sure the final concentration of salt or
detergent in the sample is as described
on page 9. You may need to manually
increase the run time for high salt or
detergent samples to obtain optimal
results.
A1 tip not aligned
Be sure to align the A1 tip properly prior
to automated loading of E-PAGE
™
96
Gel
(page 18).
Expired gel used
Use properly stored gels before the
specified expiration date.
Over-run the gel or
need more time to run
gel
Accidentally selected an incorrect
program
Select program EP for E-PAGE
™
Gels.
If you accidentally selected an incorrect
program and are at the beginning of the
run, stop the run and select the desired
program.
If you are well into the run, check the gel
to see where the loading dye is running.
Estimate the amount of time remaining
and then manually stop the run.
Protein bands
distorted on
membrane after semi-
dry blotting
Non-uniform electric field created
around wells
Be sure that the E-PAGE
™
Blotting Pad is
used correctly.
Incorrect gel orientation
Be sure that the well side of the gel is not
facing the membrane.
Weak transfer of high
molecular weight
samples during semi-
dry blotting
Not enough SDS in sample
Reduce methanol concentration in
transfer buffer from 15% to 10% if
transferring E-PAGE
48.
Make sure transfer buffer contains no
methanol if transferring E-PAGE
96.
Weak transfer of low
molecular weight
samples
Use of large pore membranes allow
small proteins to “blow through”
Use 0.2 μm nitrocellulose membrane for
optimal capture of small proteins.
Uneven transfer of
proteins and edge
lanes during semi-dry
blotting of E-PAGE
48
Gel
No methanol in transfer buffer
Use 10–15% methanol in the transfer
buffer.
Weak transfer of
proteins during semi-
wet blotting
No methanol in transfer buffer
Use 10% methanol in the transfer buffer.
Summary of Contents for E-PAGE Gels
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