Bio-Rad PROTEAN® IEF Cell Accessories User Manual

Page 38

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Place the IPG strip gel side down into a tray channel. Wet the strip by sliding it through
the rehydration solution as you place it into the channel of the tray. Minimize the amount
of solution on top of the strip and insure that each strip is completely wetted to prevent
uneven rehydration.

a. Rehydration/equilibration tray

b. Focusing tray.

Fig. 8. IPG strip placement in tray.

Apply mineral oil or similar overlay material to each channel containing an IPG Strip.
Make sure the entire strip is covered.

Place the cover lid on the tray.

Place the tray on the Peltier platform. For active rehydration be sure to align the
electrodes of the focusing tray with the Peltier electrode connections on the platform as
shown in Figure 13.

Program or select the desired method (refer to Section 2.2) and start the run. The
program may include a pause step after rehydration in order to transfer strips from the
rehydration tray to the focusing tray and/or to add electrode wicks before the
isoelectric focusing begins (or to add mineral oil).

Note: Electrode wicks should never be used during the rehydration step as the wicks can
absorb the applied sample.

10.2.2. Isoelectric Focusing

Once the IPG strips are rehydrated they can be focused with or without the use of pre-wet-
ted electrode wicks. Electrode wicks are recommended.

Prewet electrode wicks in deionized water blotting off excess water. Insert electrode wicks
directly on top of both the cathode and anode electrode wires in the focusing tray just prior
to focusing.

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