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Ettan IPGphor 3 Instrument User Manual 11-0034-58 Edition AB
4
Finally, lower the cathodic (square) end of
the strip into the channel, making sure
that the IPG gel is in contact with the strip
holder electrodes at each end. The gel can
be visually identified once the rehydration
solution begins to dye the gel. Be careful
not to trap bubbles under the strip.
Applying Immobiline DryStrip Cover Fluid
Apply Immobiline DryStrip Cover Fluid to minimize evaporation and urea
crystallization.
1
Pipet the cover fluid dropwise into one end of the strip holder until one half of
the strip is covered.
2
Then pipet the cover fluid dropwise into the other end of the strip holder,
adding fluid until the entire IPG strip is covered.
Placing the cover
Position the cover on the holder. Pressure blocks on the underside of the cover
ensure that the strip maintains good contact with the electrodes as the gel
rehydrates.
Rehydration proceeding
Rehydration can proceed on the bench top or on the Ettan IPGphor 3 unit
platform. Ensure that the holder is on a level surface. A minimum of 10 h is
required for rehydration; overnight is recommended.
Alternatively, the rehydration period can be programmed as the first step of an
Ettan IPGphor 3 protocol. This is especially convenient if temperature control
during rehydration is a concern, or if a low voltage is applied during rehydration.
3.3.2
Optional: Sample rehydration under voltage
If desired, the sample can be absorbed by the IPG strip under low voltage. This
may improve the uptake of high molecular weight proteins.
To perform rehydration under voltage:
1
Set the rehydration time to 0:00 and program Step 1 of the protocol for low
voltage (30 –100 V) for 10–12 h.
2
Program additional steps as desired to achieve complete focusing.
Operating instructions
3
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