Hoefer TE70X - Manual (Page 24)

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p16
problem solution
Inefficient binding to membrane
Chemical parameters
Prepare transfer buffer without SDS. (SDS can
improve transfer efficiency but can interfere with
protein binding to a PVDF membrane.)
Add 10 – 20% methanol to the transfer buffer to
enhance binding to nitrocellulose or PVDF.
Membrane parameters Use a membrane with a smaller pore size (0.20 µm) if
proteins pass through the membrane.
Place a membrane both over and under the gel to
capture any proteins migrating in the opposite direction.
Check if too much sample is available for the binding
surface area by placing two membranes instead of one.
If “blow through” occurs, reduce the sample load.
Wear gloves when handling membranes.
Store membranes at ambient temperature and out of
direct sunlight.
Check shelf life of nitrocellulose membrane, replace
if necessary.
TE70XP, TE77XP
Current reading
Maximum voltage (30 V) has been reached.
lower than setting Buffer may be depleted.
No output current Transfer stack is not making contact with the upper
electrode. Add buffer soaked blot papers.
“dry” error message Instrument has detected large changes in transfer
stack resistance.
Add more buffer soaked blot papers in future transfers.
No output voltage Transfer stack is being short circuited.
Note: For more troubleshooting
ideas, refer to Bjerrum, O.J.
et al. (1988).