GE Ettan IPGphor 3 Operating Instructions Manual page 52

5 Operation
5.1 Preparations before start
5.1.1 Use the strip holders
Apply Immobiline DryStrip
cover fluid
Follow the instructions below to apply Immobiline DryStrip Cover Fluid to minimize
evaporation and urea crystallization.
Step
1
2
Place the strip holder 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 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 re-
hydration is a concern, or if a low voltage is applied during rehydration.
Sample rehydration under
voltage (optional)
Follow the instructions below to perform 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.
Step
1
2
52
Action
Pipet the cover fluid dropwise into one end of the strip holder until one half
of the strip is covered.
Later, pipet the cover fluid dropwise into the other end of the strip holder,
adding fluid until the entire IPG strip is covered.
Action
Set the rehydration time to 0:00 and program Step 1 of the protocol for low
voltage (30 to 100 V) for 10 to12 h, see Edit step or gradient, on page 79
Program additional steps as desired to achieve complete focusing.
Ettan IPGphor 3 Operating Instructions 28-9635-51 AC